oEPA
           United States
           Environmental Protection
           Agency
            Office of Research and
            Development
            Washington DC 20460
EPA/600/6-88/004
February 1988
Reference Physiological
Parameters in
Pharmacokinetic Modeling

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                                             EPA/600/6-88/004
                                             February 1988
Reference Physiological Parameters in Pharmacokinetic Modeling
             Angela D. Arms and Curtis C. Travis
               Oak Ridge National Laboratory
              Oak Ridge, Tennessee 37831-6109
        Office of Health and Environmental Assessment
             Office of Research and Development
            U.S. Environmental Protection Agency
                  Washington, B.C. 20460
                                              Printed on Recycled Paper

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                                DISCLAIMER







     This document has been reviewed in accordance with  U.S.  Environmental



Protection Agency policy and approved for publication.  Mention of  trade



names or commercial products does not constitute endorsement or recommendation



for use.

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                                CONTENTS
Tables	

Figures	•

Abstract	

Authors, Contributors,  and Reviewers	      iz

1.  SUMMARY	     1-1

2.  INTRODUCTION	     2-1

3.  PHARMACOKINETIC MODELING	     3-1

    3.1.  DESCRIPTION	     3-1

4.  PHYSIOLOGICAL PARAMETERS	     4-1

    4.1.  BODY WEIGHTS	     4-2

          4.1.1.  Reference Values  	     4-2
          4.1.2.  Documentation 	     4-2

    4.2.  TISSUE VOLUMES	     4-4

          4.2.1.  Reference Values	     4-4
          4.2.2.  Documentation 	     4-5
          4.2.3.  Values Used in Pharaaco kino tic Models	     4-9

    4.3.  CARDIAC OUTPUT	    4-17

          4.3.1.  Reference Values	    4-18
          4.3.2.  Documentation 	   4-18
          4.3.3.  Values Used in Pharaacokinetic Models	   4-29

    4.4.  CARDIAC OUTPUT DISTRIBUTION	   4-30

          4.4.1.  Reference Values	   4-30
          4.4.2.  Documentation 	   4-30
          4.4.3.  Values Used in Pharmacokinetic Models	   4-33
                                    iii

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                           CONTENTS  (continued)



                                                                       Page

    4.5.  RESPIRATION PARAMETERS	    4-41

          4.5.1.  Reference Values	    4-42
          4.5.2.  Documentation	    4-43
          4.5.3.  Values Used in Pharmacokine tic  Models	    4-61

5.  SCALING	     5-1

    5.1.  CARDIAC OUTPUT	     5-2

          5.1.1.  Mice	     5-3
          5.1.2.  Rats	     5-4
          5.1.3.  Humans 	     5-6

    5.2.  MINUTE VENTILATION	     5-7

          5.2.1.  Mice	     5-7
          5.2.2.  Rats	     5-9
          5.2.3.  Humans 	     5-13

6.  APPENDIX A: TABLE OF PARTITION COEFFICIENTS	     6-1

7.  REFERENCES	     7-1
                                     iv

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                                  TABLES
                                                                        Page

1-1      Reference physiological parameters	      1~1

3-1      Abbreviations and symbols used in describing
         a pharma co kinetic model	.	      3-4

4-1      Physiological parameters used for modeling	      4-1

4-2      Reference body weights of mice,  rats,  and humans	      4-2

4-3      Reference tissue volumes of mice, rats,  and humans  ....      4-4

4-4      Reference cardiac output of mice, rats,  and
         humans	     4-18

4-5      Absolute cardiac output of mice	     4-19

4-6      Relative cardiac output of mice	     4-20

4-7      Absolute cardiac output of unanesthetized rats  	     4-21

4-8      Absolute cardiac output of anesthetized rats	     4-22

4-9      Relative cardiac output of unanesthetized rats  	     4-25

4-10     Relative cardiac output of anesthetized rats  	     4-26

4-11     Absolute cardiac output of humans 	     4-27

4-12     Cardiac output values used in ph arm a co kinetic models  ". . .     4-29

4-13     Reference tissue perfusion rates of mice, rats,
         and humans	     4-30

4-14     Reference respiration parameters of mice, rats,
         and humans	«,    4-43

4-15     Absolute minute volumes of unanesthetized mice	    4-44

4-16     Absolute minute volumes of anesthetized mice	    4-45

4-17     Relative minute volumes of unane sthetized mice	    4-46

4-18     Relative minute volumes of anesthetized mice	    4-46

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TABLES (continued)

4-19
4-20
4-21
4-22
4-23
4-24
4-25
4-26
4-27
4-28
4-29
4-30

4-31
5-1

5-2
5-3
5-4
5-5
5-6

5-7
5-8
5-9
5-10
5-11

Respiratory frequencies of unane sthetized mice. . .
Respiratory frequencies of anesthetized mice 	
Absolute minute volumes of unane sthetized rats 	 „
Absolute minute volumes of anesthetized rats 	
Relative minute volumes of unane sthetized rats 	


Respiratory frequencies of anesthetized rats 	




if
Alveolar ventilation used in pharmaco kinetic models ....
Constants for calculating cardiac output



Scaling cardiac output of anesthetized rats 	

Constants for calculating minute volume

Sealing minute volume of unane sthetized mice 	
Scaling minute volume of anesthetized mice 	
Scaling minute volume of unane sthetized rats 	
Scaling minute volume of anesthetized rats 	
Scaling minute volumes of humans ......•••••• «
Page
4-47
4-48
4-49
4-50
4-51
4-52
4-55
4-56
4-57
4-58
4-59
4-61

4-62

5-3
5-3
5-4
5-5
5-6

5-7
5-8
5-8
5-10
5-12
5-13
         vi

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                                  FIGURES
                                                                         Page

3-1      Diagram of a typical pharmaco kinetic model used to  simulate
         the behavior of inhaled volatile organics	     3-3
                                    vii

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                                 AB SIS ACT







     This document presents a compilation  of  measured value s for




physiological parameters used in pharmacokinetic  modeling.  The




physiological parameters include body weight,  tissue volumes,  cardiac




output, cardiac output distribution, and the respiration parameters.




Reference values for use in risk assessment are given for each of the




physiological parameters based on analyses of valid measurements obtained




from the literature and other reliable sources. The proposed reference




values are for generic mice and rats without regard to sex or strain.




Reference values for humans are without regard to age or sex. Differences




between the  sexes in mice, rats, and humans are accounted for by scaling




the reference parameters within  species on the basis of body weight.




Reference physiological parameters  are for a 0.025 kg mouse, 0.25 kg rat,




and a 70 kg man.
                                   viii

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                   AUTHORS,  CONTRIBUTORS,  AND REVIEWERS
AUTHORS

Angela D. Arms
Office of Risk Analysis
Health and Safety Division
Oak Ridge National Laboratory
Oak Ridge, TN  37831-6109
Curtis C. Travis
Office of Risk Analysis
Health and Safety Division
Oak Ridge National Laboratory
Oak Ridge, TN  37831-6109
CONTRIBUTORS

Janis G. Pruett                                         APPENDIX A
Information, Research, and
Analysis Department
Biology Division
Oak Ridge National Laboratory
Oak Ridge,  TN  37831
REVIEWERS

The following individuals provided
peer review of this document and/or
earlier drafts of this document.
Jerry N. Blancato
Office of Research and Development
U. S. Environmental Protection Agency
Washington, D. C.  20460
Chao W. Chen
Office of Research and Development
U. S. Environmental Protection Agency
Washington, D. C.  20460
                                     iz

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 Vera Fiserova-Bergerova Thomas
 Department of Anesthesiology
 University of Miami School of Medicine
 Miami, FL  33101
 Joe L. Mauderly
 Inhalation Toxicology Research Institute
 Lovelace Biomedical  and Environmental
 Research Institute,  Inc.
 Albuquerque,  NM  87185
 Fred J.  Miller
 Health Effects Research  Laboratory
 U.  S.  Environmental  Protection Agency
 Research Triangle  Park,  NC  27711
Ellen J. O'Flaherty
Institute of Environmental Health
lettering Laboratory
University of Cincinnati Medical Center
Cincinnati, OH  45267-0056
Richard W. Walentovicz
Office of Research and Development
U. S. Environmental Protection Agency
Washington, D. C.  20460

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                                1.  SUMMARY



Table 1-1 summarizes the recommended reference physiological parameters for

use in pharmacokinetic modeling.
0.055
0.10
0.05
0.70
0.04
0.07
0.05
0.75
0.026
0.19
0.05
0.62
               TABLE 1-1. REFERENCE PHYSIOLOGICAL PARAMETERS


                                      Mouse    Rat    Human

               Body weights (kg)      0.025   0.25    70.0

               Tissue volumes
               (fractions)

                  Liver
                  Fat
                  vac
                  m

               Cardiac output         0.017   0.083    6.2
               (1/min)

               Tissue perfusion
               (fractions)

                  Liver
                  Fat
                  VfiG
                  m

               Minute volume          0.037   0.174    7.5
               (1/min)

               Alveolar ventilation   0.025   0.117    5.0
               (1/min)
0.25
0.09
0.51
0.15
0.25
0.09
0.51
0.15
0.26
0.05
0.44
0.25
                                     1-1

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                             2.  INTRODUCTION







     Pharmaco kine tics is the science of quantitatively predicting  the  fate




of an exogenous substance in an organism.  Utilizing computational




techniques,  pharmacokinetics provides the means of  studying the uptake,




distribution, metabolism, and excretion of chemicals by the body.  This  is




accomplished by dividing the body  into various anatomical compartments.




The mathematical representation of these compartments provides a




description of the  time  course of  drug disposition  throughout the  body.




Pharmacokine tics eliminates some of  the ambiguities in determining the risk




of human exposure  to environmental chemicals and provides a basis  for




evaluating the  scientific assumptions upon which the risk assessment




process is based.




     A recent development in the area of pharmacokine tics is the advent  of




physiologically-based pharmaco kine tic (PBPK) models.  Relying on actual




physiological parameters such  as body weight, breathing rates, cardiac




output, blood flow  rates, tissue volumes, etc., to  describe the metabolic




process, the PBPK models can relate  exposure concentrations to organ




concentrations  over  a range of exposure  conditions.  This report provides  a




literature review  of the physiological parameters  used in PBPK models, and




recommends reference physiological parameters for  use in risk assessment.
                                     2-1

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                       3.   PHARMACOKINETIC MODELING






3.1. DESCRIPTION






     A pharmaco kinetic model is a set of equations that can be used to




describe the time course of a parent chemical or metabolite in an animal




system.  There are two types of pharmacokinetic models: data-based and




physiologically-based (NEC, 1986).  A data-based mode 1 divides the animal




system into a  series of compartments which, in general, do not represent




real,  identifiable anatomic regions of the body.  In applying these models,




time-course concentration  curves are first determined from in vivo animal




experiments.  Then, model  compartment volumes and rate constants are




determined by  trial and error so that the model predictions lit the




empirical data.  These models are useful for interpolation and limited




extrapolation within the same species.  However, since the parameters in




these  data-based model s generally do not correspond to physiologically-




identifiable entities, they do not allow for extrapolation across animal




species.




     A physiologically-based pharmaco kinetic model is  comprised of a series




of compartments representing organs or tissue groups with realistic weights




and blood flows.  These models require a variety of physiological




information: tissue volumes, blood flow rates to tissues, cardiac output,




alveolar ventilation rates  (for volatile compounds) and, possibly, membrane




permeabilities.  The models also utilize biochemical  information  such as




air/blood partition coefficients, and metabolic parameters.   The  uniqueness




of the physiologically-based approach is this reliance on measured




physiological and biochemical parameters.  An appealing  aspect  of  these




physiological models is that they allow ready extrapolation  of  observed
                                    3-1

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experimental results from a test species to an untested species simply by




placing the appropriate physiological and biochemical parameters in the




model.  Similarly, the effect of route of administration can be




investigated by allowing for different administration pathways.




     It should be pointed out that no one pharmacokine tic model can be used




to determine the distribution of all chemicals.  The number of compartments




and the way they are connected will vary from chemical to chemical




depending upon the chemical's metabolic behavior and the nature of the




questions being asked concerning dose to target tissues.




     Despite this fact, most physiologically-based pharmacokine tic models




in current use divide the body into four physiological groups, all




connected by the arterial and venous blood flow pathways (see Figure 3-1




and Table 3-1).  The first group is the vessel-rich group (VUG) and is made




up of those tissues most profusely supplied with bloodvessels.  These




include the brain, heart, kidney, and viscera.  The second group is




composed of muscle and skin and is called the muscle group, (MB).  The




third group is composed of adipose (fat) tissue.  The fourth  group contains




organs with a high capacity to metabolize (principally  liver). , Each  tissue




group is described mathematically by a set of differential equations which




calculate the rate of change of  the amount of chemical  in each compartment.




Metabolism, occurring chiefly in the liver,  is  described by a combination




of a linear metabolic component  and a Michael is-Menten  component accounting




for saturable metabolism.  Again, we  stress  that  other  model  descriptions




are possible,  but they will, in  general, have  the  same  physiological




parameters.
                                     3-2

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                                              ORNL-OWG H42MR
o.,.
^inh
°> t
c... "-
C.f
C,m
C,,
c,,
ALVEOLAR
SPACE
LUNG
BLOOD

FAT
TlCCl 1C
GROUP

MUSCLE
GROUP

VESSEL
Q1/*U
GROUP

LIVER
METABOLIZING
TISSUE
GROUP
Q.,v
c... *
Qb
c.,,
Of
C.r,
t Qm
1 c.rl
Or
' c.rl
Q,
" c.rl
                        METABOLITES
                      (LINEAR PATHWAY)
                              METABOLITES
                           (MICHAELIS-MENTEN)
FIGURE 3-1.
Ditfraa of  i  typical  pbmacokinetic
•odel used  to «i«ultte  the behavior
of inhaled  volatile  organic*.  The
•odel divides the  body  into four
physiological  groups,  all connected
by blood flow pathways.   The symbols
are defined in Table  3-1.
                             3-3

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                TABLE 3-1. NOMENCLATURE USED IN DESCRIBING A
                           PHYSIOLOGICALLY-BASED PHARMAOOKINETIC MODEL


        Alveolar ventilation rate  (liters air/or)

        Concentration in  inhaled air  (mg/liter air)

Calv    Concentration in  alveolar  air  (mg/liter air)

Xb      Blood/air  partition coefficient  (liters air/liters blood)

Qb      Cardiac output  (liters blood/hr)

Cart    Concentration in  arterial  blood  (mg/liter blood)

Cyen    Concentration in  mixed venous  blood  (mg/liter blood)

Vmax    Michaelis-Menten  metabolism rate  (mg/hr)

Km      Michael is  constant  (mg/liter  blood)

Kf     Linear metabolism rate  (hr~l)

Am     Amount metabolized  in the  liver  (mg)

Qi     Blood flow rate to  tissue  group  i  (liters blood/hr)*

Vi     Volume of  tissue  group  i  (liters)

Ci     Concentration in  tissue  group  i  (mg/liter)

Ai     Amount in  tissue  group  i  (mg)

Cvi    Concentration in  venous  blood leaving tissue group  i  (mg/liter blood)

Xi     Tissue/blood partition  coefficient for tissue i  (liters  blood/liter  i)

Xi/a   Tissue/air partition coefficient for tissue  i (liters air/liter  i)

k      Gavage or  oral rate constant  (hr~l)

DO     Total quantity of PCE absorbed via gavage route  (mg)

"Subscripts (i) for tissue groups or  compartments:


     1   Liver (metabolizing tissue  group)

     f   Fat tissue group

     r   Vessel-rich tissue group

     m   Muscle tissue group
                                          3-4

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                       4.    PHYSIOLOGICAL  PARAMETERS


     The physiological parameters typically  used  in pharmacokine tic

modeling are listed below.
                    TABLE 4-1.  PHYSIOLOGICAL  PARAMETERS
                               USED FOR MD EEL ING.
                    PARAMETERS
                    Body weight (kg)
                    Cardiac output (1/min)
                    Minute volume (1/min)
                    Alveolar ventilation (1/min)
                    Physiological dead space  (%)
                   'Frequency (breaths/min)
                    ORGANS
                    (Volumes and Blood Flows)

                    Liver
                    Fat
                    Vessel-Rich Group
                    Muscle Group
Measured values of these parameters in mice and rats are age,  sex,  and

strain-dependent.  For example, female rats tend to have higher mass-

specific ventilation rates than males, and young rats have values higher

than mature rats.  In addition, the status of the animals during

measurement (body position, conditioning, etc.,) and the measurement

technique can have substantial influences.  Lack of data for many

physiological parameters, however, limits attempts to account for  these

factors.  Our reference parameters are for a generic mouse  or rat, without

regard to sex or strain.  Differences between  sexes are  accounted  for  by
                                    4-1

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scaling the reference  parameters within species on  the basis of body




weight.  Reference  physiological values for  humans  are for a resting 70 kg




man.  For rodents,  the reference physiological parameters are for a 0.025




kg mouse and a 0.25 kg rat at rest.




     We will now review the measured values  of these  parameters found  in




the literature.   Each  section is organized as follows: first, a summary




table of the recommended reference  values is presented;  then  a literature




review  is undertaken to support the recommended reference values; and




finally, a summary of  the actual values of parameters used  in various




ph arm a co kinetic models is presented.









4.1. BOD7 WEIGHTS






4.1.1.  Reference Values









                     TABLE 4-2. REFERENCE BODY WEIGHTS






                       Mice       Rats        Human •




                     0.025 kg    0.25 kg     70.0 kg
4.1.2.  Documentation of Reference Values
                                   Mice
     The U.S. Environmental Protection Agency (1980) recommends a  standard




body weight of 0.03 kg for an adult mouse.  Other reported standards  in the




literature include 0.035 kg (ARS/Spr ague-Daw ley, 1974), 0.023 kg




(Bozenbanm, 1982) and 0.02 kg (Lehman, 1959).  Altnan and Dittmer  (1972)
                                    4-2

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provided growth data on seven strains of mice from birth to about six




months of age, by which time their body weights were considered stable.




The mean terminal body weight for both sexes of all strains combined was




0.032 kg (Durkin, 1985).  Therefore,  the reference value of 0.03 kg




recommended by the EPA seems reasonable.   However, most pharmacokine tic




studies are performed on mice before they reach terminal body weight.   For




the purposes of pharmacokine tic modeling, we are recommending a reference




body weight of 0.025 kg.






                                   Rats






     The U.S. EPA (1980) recommends a standard body weight of 0.35 kg for




an adult rat.  Other reported standards in the literature were 0.45 kg




(ARS/Spr ague -Daw ley, 1974), 0.25 kg (Boxenbaum, 1982) and 0.40 kg (Lehman,




1959).  Growth data for two strains of rats were provided by Altaian and




Dittmer (1972),  in which the mean terminal body weight for both  sexes and




strains combined was 0.334 kg (Durkin, 1985).  Thus, the reference value of




0.35 kg for rats proposed by the EPA seems justifiable; however, most




pharmacokine tic studies are performed on rats before they reach  terminal




body weight.  For the purposes of pharmacokine tic modeling, we are




recommending a reference body weight of 0.25 kg.






                                  Humans






     The International Commission on Radiological Protection  (ICRP)




recommends a body weight of 70 kg for an adult reference man  (Snyder  et




al.,  1975).  This value is derived from  the  total body  weight  of organs,




tissues,  and components assumed  to include blood  vessels  and  other  body




fluids.  The U.S. EPA  (1980) concurs with  the  ICRP  (1975)  in recommending
                                    4-3

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70 kg as the acceptable reference  value  for an adult man.









4 .2 . TISSUE VCLUJES






     Tissue volumes cited in pharmacokine tic models generally reference  the




power functions reported by Adolph (1949).   When these functions are




applied to mice and rats, they are only  approximately correct.  It seems




prudent to base tissue volumes on direct measurements when possible;




however, they can be determined indirectly through optimization of the




appropriate parameters in a pharmacokine tic model.









4.2.1.  Reference Values






     Reference tissue volumes are presented in Table 4-3 as a percentage of




total body weight.  Tissue volumes in liters may be obtained by multiplying




total body weight  (section 4.1.) by the tissue volume fraction and assuming




that one kilogram of tissue has a volume of one liter.









                TABLE 4-3. REFERENCE TISSUE VCLUJE FRACTIONS
                          Mouse         Rat         Human




                Liver      0.055        0.04        0.026




                Fat        0.10         0.07        0.19




                VBG        0.05         0.05        0.05




                1C         0.70         0.75        0.62
                                     4-4

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4.2.2.  Documentation of Reference Values




Data indicate that the liver grows steadily  throughout  life  and remains  in


a fairly constant relationship to body weight.   Liver weight/body weight


ratios appear to be highly strain-dependent.
Liver
     Liver weight/body weight ratios in mice  are  more  variable  than  in


rats, ranging from 3.5 to 8.0%,  depending on strain.   Specter  (1956)


recorded liver volume measurements of 5.63% of  body  weight  for  the jumping


mouse and 4.56% of body weight for the meadow mouse.   Kurnick and Kernen


(1962) reported that after weaning (10 weeks),  liver weight maintained  a


uniform relationship to body weight (4.5 to 5.8%)  in mice.   Bischoff et al .


(1971) reported a value of 6.0%, while Tuey and Matthews (1980)  found a


value of 5.8%.  On the basis of  limited data,  Crispens (1975) reported


values of about 3.5%.  Gellatly  (1975) reported the  liver weight of  mice on
                                                             •

a stock pelleted diet (standard diet) as 5.64% of body weight  for males and


5.44% for females.


     Fieri et al. (1980) found male Balb/c mice at 2 months of  age  had  a


liver weight/body weight ratio of 4.8%, compared to a 4.4 to 4.5% range at


12 to 24 months.  Kj el lit rand et al.  (1981) reported a liver weight/body


weight range of 3.6 to 5.2% in control NIRI mice weighing 0.027 to 0.036


kg.   Ruebner et al.  (1984) reported  liver weight/body weight ratios in


three strains of control mice.  In male C57B1/6 mice,  43 to 132-weeks old,


the  liver weight/body weight range was 4.7 to 7.7%.  The geometric mean of


the  values was 5.68%.  In male C3H/He mice, 41 to Ill-weeks old, the liver
                                    4-5

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weight/body weight range was 4.3  to 8.0%.   The  geometric mean of the values




was 5.48%. In male B6C3Fi mice,  21 to 119-weeks old,  the liver weight/body




weight range was 4.0 to 7.3%.  The geometric mean of  the values was  5.6%.




De Marte and Enesco (1986) reported a liver weight/body weight ratio of




5.7% for eight-week old male Swiss Albino  mice  weighing 0.034 kg.  For mice




24 to 78-weeks old, the percentage remained fairly constant  from 4.2 to




4.7%.




     The above data generally indicate that young mice  have  liver




weight/body weight ratios higher than older mice.  Based on body  weight,




oar reference mouse of 0.025 kg would qualify as a young mouse.   Therefore,




the reference liver volume of 5.5% of total body weight is a reasonable




one.




     Jackson  (1913) found that in the albino rat the liver was  on the




average 4.42% of  total body weight at one  year of age.   Webster  et al.




(1947) determined  liver weight/body weight ratios in 500 Albino  rats with




no consideration  given to age.  Rats weighing 0.250 kg gave  a liver




weight/body weight ratio of 4.09% and 0.300 kg rats,  4*.0%.  Addis and Gray




(1950)  found  a range of 3.0  to 6.0%  in Albino rats; the 6.0% value was for




young rats.   No values higher than 3.8% were found for rats weighing more




than 0.200 kg.  De Lector-Israel (1971) cited a liver weight/body weight




ratio of 3.2% for  control rats 18 to 24-months old and 3.1% for control




rats 24 to 27-month s old.  Coniglio  et al.  (1979) studied the liver




weight/body weight ratio as  a function of  age  in male  Fischer rats.  They




found body weight  reached its peak at 12 months,  then  declined steadily




from 12 to 30 months.  From  six to 20 months,  the liver weight/body weight




ratio remained fairly constant at 3.1 to 3.5%.   For  a  young rat 2 to 4-




months old, the liver weight/body weight ratio was 3.8%.  Kj el 1st rand et
                                    4-6

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al. (1981) reported a liver weight/body weight ratio of 3.89 to 4.29% in




control Spr ague-Daw ley rats weighing 0.200 to 0.394 kg.  Van Bezooijen




(1984) reported liver weight/body weight ratio as a function of age in six




different strains of rats.  From one to 27 months,  the male Fischer 344 rat




gave a liver weight/body weight ratio of 2.9 to 2.7%,  declining with age.




Similar findings were reported for both the male and female Wistar rat and




the male and female WAG/Rij rats.  Male Spr ague-Daw ley rats also showed a




decline in the liver weight/body weight ratio with a range of 4.5 to 3.4%




from three to 16 months of age.  A measured liver volume percentage of 4.0%




of body weight was reported by Lutz et al. (1977) and a range of 4.2 to




6.3% by Jansky and Hart (1968).




     The above data indicate that liver weight/body weight ratios in rats




decline with age.  Based on body weight, our reference rat of 0.250 kg




would qualify as a young rat.  Therefore, the reference liver volume of




4.0% of total body weight  is a reasonable one.




     A liver volume fraction of 2.6% in the adult reference man is recorded




in ICRP (1975; Boyd, 1941).  Sherlock  (1968) reported  the liver comprised




2.0% of the adult body weight.  DeLand and North (1968) found a linear




relationship exists between liver weight and body weight.  They reported a




liver weight volume of 2.2% in a 70 kg man.  We are recommending a




reference human liver volume of 2.6%.






                                Fat VolT""e






     The recommended fat volume fraction for mice  is  supported  by  measured




values of 9.8% (Tuey and Matthew*, 1980). and an average  of  10.7%  (range  of




8.3 to 15.5%) for four different  strains of mice  (Dawson,  1970).   Dawson




(1970) found that body weight was not  a  good predictor of  fat  content in
                                    4-7

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the four strains of mice.   We recommend a reference fat volume percentage




for mice of 10%.




     An empirical determination of 7.0% of the body fat fraction of rats




was reported by Caster et al. (1956)and Lutz et al. (1977).  Reed et al.




(1930), feeding rats special diets rich in fats and carbohydrates, reported




an average value of 8.8%.  The fat volume percentage in humans varies from




11.5 to 36% in males and 15 to 38% in females.  We follow Reference Man and




recommend a fat volume percentage of 19.0%.






                            VRG and BIG Volumes






     The VRG and muscle volumes are not necessarily supported by  eapirical




measurements since these compartments are difficult to identify.  The VRG




as defined by one  investigator may include or delete organs considered  in




another's definition of VEG.  In order to be exact, it becomes necessary to




summate measured values from the individual organs which comprise  the VRG.




For example, Ramsey and Andersen  (1984) include the brain, heart, kidney.




and viscera in  their definition of VRG.  To obtain an accurate measured




value  for the VRG, one would have to sumaate  the  individual measurement s of




the four organs to recommend a total value for the VRG.




     Our definition of 1C  includes the muscle and skin.  Skeletal muscle




represents 40% of  total body weight.  The weight  of this tissue  includes




the connective  tissue, bloodvessels, blood,  lymph, etc.,  which  is normally




associated with skeletal muscle  (ICEP, 1975).  Our  reference  muscle volume




value  of 62% also  includes  smooth muscle.   Skin accounts  for  3.7% of total




body weight in a 70 kg man.
                                    4-8

-------
4.2.3.  Values Used in Pharmacokinetic Models


     Since direct measurement of many of the physiological parameters used

in pharmacokine tic models is difficult,  many investigators believe that the

best determination of the parameters results from optimization.   It is

important to clarify that the physiological parameters used in these  model

simulations produce the best fit to experimental data and may not represent

physiological values for a reference man.  We now review the combination of

empirically-determined and optimized parameters for tissue volumes which

have appeared in the literature.  The values are given as a percentage of

total body weight for each species.



                                   Mice


     Bischoff et al. (1971) used tissue volumes verified from laboratory

measurements.  The tissue volumes for a 0.022 kg mouse were obtained for

six compartments:



                            Tissue	Percent

                            Blood          7.6
                            Muscle       45.0
                            Liver          5.9
                            Kidney         1.5
                            61  tract       6.8
                            Gut lumen      6.8


The gastrointestinal tract and  the  gut-lumen were  the  two  tissue  groups  not

independently measured.  Total  tissue volume accounted for  approximately

74% of body weight.

     Dedrick et al. (1973) based most  of  his tissue  volumes for  a 0.022 kg

mouse on the values reported by Bischoff  et al.  (1971).
                                    4-9

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                            Tissue	Pe r co nt

                            Blood         7.6
                            Lean         45.0
                            Liver         5.9
                            Kidney        1.5
                            Heart         0.43
                            Gut lumen     6.8
                            Marrow        2.7
Unlike Bischoff (1971). Dedrick did not include a separate compartment for

the gastrointestinal tract.   Marrow tissue represented approximately 3.0%

of body weight and heart tissue 0.43%.  Total tissue volume accounted for

70% of total body weight.  Neither Bischoff (1971)  or Dedrick et al. (1973)

recognized fat as a tissue compartment.

     Tney and Matthews (1980) provided physiological values for a 0.038 kg

mouse, using the same tissue volumes for the blood,  muscle, and gut lumen

compartments as Dedrick et al.  (1973).  The average  values for the  skin and

fat compartments were 14.5 and 9.8% of body weight,  respectively,  based on

measurements obtained from complete dissection of mice in their laboratory.

Total tissue .volume accounted for 89% of the total body weight of the

mouse.

     Ramsey and Andersen (1984) used the physiological values for a 0.022

kg mouse  suggested by previous investigators with one important exception.

They included fat as a tissue with a volume representing  9.0% of body

weight.



                         Tissue	Percent

                         Liver               4.0
                         Fat                 9.0
                         Richly perfused     5.0
                         Muscle             73.0
                                   4-10

-------
The  SUB of  the  tissue volumes accounted for 91% of  total body weight  of  the

mouse, with  the remaining 9.0% designated as  skeletal  tissue.  Unlike

previous  investigators. Ramsey and Andersen  (1984)  did not  specify  a  blood

volume, since  the blood did not act as a  significant repository  for  the

chemical  under  investigation.

     Andersen et al.  (1987) reported the following  physiological values  for

a 0.0345  kg mouse:



                        Ti ssue	Per cent

                        Liver                 4.0
                        Fat                   4.0
                        Rapidly perfused      5.0
                        Slowly perfused     78.0



     In summary, values used in pharmacokinetic modeling for mice  tissue

volumes varied  from 4.0 to 6.0% for the liver, 4.0  to  9.0%  for fat,  50%  for

VRG  and 73  to 78% for  the muscle  group.



                                   Rats


     Bischoff et al.  (1971) provided tissue volumes for  a 0.200  kg rat

based on  his previous work and the correlations of  Adolph  (1949):



                            Ti ssue	Pe r ce nt

                            Blood        7.5
                            Muscle        50.0
                            Kidney        0.95
                            Liver        4.15
                            GI tract      5.5
                            Gut lumen     5.5


Tissue volumes  accounted for approximately  74%  of total  body weight.
                                    4-11

-------
Bischoff's model neglects the fat compartment.

     Harrison and Gibaldi (1977)  gave tissue  volumes for  a  0.360  kg  rat

based on measurements by Thompson and Hollis  (1958). Jansky and Hart (1968)

and the correlations of Adolph (1949):

                       Tissue             Percent
                                            4.92
                       Skeletal muscle     50.0
                       Kidneys              0.94
                       Liver                3.72
                       Heart                0.50
                       GI tissues           3.0
                       61 co nte nt s          4.8
                       Skin, fat. etca     25.0

                       a Miscellaneous compartment
                         for all other body regions


Harrison and Gibaldi (1977)  included the heart as a separate  compartment,

in addition to recognizing fat as a miscellaneous compartment for all other

body regions (skin, etc).  The sum of tissue volumes accounted for

approximately 93% of total body weight.

     Lutz et al.  (1977) gave the following tissue volumes for a 0.250 kg

rat:



                            Tissue	Percent

                            Blood         9.0
                            Muscle       50.0
                            Liver         4.0
                            Skin         16.0
                            Fat           7.0
                            Gut lumen     5.6


Lutz et al.  (1977)  based their muscle volume of 50% of total body weight on

the data of  Bischoff et al.  (1971).  Liver volume was measured;  skin and

adipose (fat) tissues represented average values obtained by laboratory

dissection of the rats.  These values are also consistent with  the
                                   4-12

-------
previously published data of Freudenberger (1933)  and Reed e t  al.  (1930).

Tissue volumes accounted for approximately 92% of  total  body weight.

     Ramsey and Andersen (1984)  reported tissue volumes  for a  0.30 kg  rat

as:



                         Tissue	Percent

                         Liver               4.0
                         Fat                 9.0
                         Richly  perfused     5.0
                         Muscle              73.0


Total tissue volume accounted for 91% of total body weight.

     Fi serova-Ber gerova and Hughes (1983) estimated the  following tissue

volumes (liver designated as a separate compartment) for a 0.250 kg rat:



                             Tissue   Percent

                             Liver      4.0
                             Fat        0.40
                             VBG        5.6
                             Muscle    72.0


Another compartment accounting for 15.4% of the total tissue volume

represented the skeleton, blood, and lungs.  The parameters for the rat

were tested in a model simulation of halothane in a 0.200 kg rat

(Fi serova-Ber gerova and Kawiecki, 1984).  The fat parameter is quite low in

comparison with other model simulations.  This can be attributed  to the

young age of the rat under  study and the desire to obtain  the best fit to

experimental data (Fi serova-Ber gerova, 1987) .

     Andersen et al. (1987) reported tissue volumes for a  0.233 kg rat:
                                   4-13

-------
                        Tissue	Pe r ce at

                        Liver                4,0
                        Fat                  7.0
                        Rapidly  perfused     5.0
                        Slowly perfused      75.0
                                  Humans


     Mapleson (1963)  used the  following tissue  percentages  in his model

simulation:



                  Tissue	Percent

                  Adrenals                          0.11
                  Kidneys                           1.52
                  Thyroid                           0.09
                  Gray matter                        1.60
                  Heart                             0.64
                  Other small  organs and  glands     0.30
                  Liver plus  portal  system          6.97
                  White matter                      1.21
                  Red marrow                         2.10
                  Muscle                            43.38
                  Skin plus nutritive shunt          4.33
                  Non-fat subcutaneous              6.91
                  Fat                              14.46


Each value listed includes the volume of  blood in equilibrim with the

tissue.   Bone cortex,  fatty marrow tissue, and arterial and venous blood

accounted for approximately 21% of total  tissue volume.

     Eger (1963)  recommended the following tissue volume  percentages for a

70 kg man:
                                   4-14

-------
                             Ti s sue   Pe r ce nt

                             VUG        8.6
                             Fat       21.0
                             Muscle    47.0
                             VPG       18.0
Eger's VBG included the brain, heart, kidneys, hepatoportal system (liver)

and the endocrine glands.  The VPG included bone and cartilage.

     Bischoff et al. (1971) provided the following tissue volumes for a 70

kg man:



                            Tissue	Pe r ce nt

                            Blood         7.15
                            Muscle       50.0
                            Kidney        0.40
                            Liver         2.0
                            GI tract      3.0
                            Gut lumen     3.0


The gut and  gut-lumen volumes were not  independently measured, but are

actually effective values.  Total percentage  of  tissue volumes represented

66% of body  weight for  a 70 kg man.  Bischoff  neglects the  fat compartment

for man.

     The tissue volumes  cited by Dedrick et al.  (1972) were essentially

taken from the data of  Mapleson  (1963):



                             Tissue    Percent
Blood
Liver
Gut
Heart
Kidney
Lean
Marrow
3.8
2.4
4.5
0.64
1.5
38.6
2.8
                                    4-15

-------
     Each values listed includes the  volume  of  blood  in equilibrium with

the tissue.   Calculation of  the  liver,  and gat  values were  derived from

Mapleson (1963) as well as other sources.   The  muscle value was lowered  to

38.6% from the original value  of 58.7%  to  achieve  an  adequate  simulation of

the data (Dedricket al.,  1972).

     Fiserova-Bergerova et al.  (1984) estimated the following  tissue

volumes for a 70 kg man at rest:



                            Tissue   Percent

                            Liver      4.0
                            Fat       15.0
                            VBB        5.0
                            Muscle     49.0


The VEG included brain, gastrointestinal tract,  glands,  heart,  kidneys,  and

spleen.   The muscle group included muscles and skin,  and the  fat

compartment included adipose tissue  and white  marrow.

     Ramsey and Andersen (1984)  provided the following  tissue  volumes for

an 83 kg man:



                         Tissue	    Percent

                         Liver               4.0
                         Fat                 9.0
                         Richly  perfused     5.0
                         Muscle              73.0


These tissue volumes were assumed to be proportional  to body weight, and

were scaled up in direct proportion to the ratio of  body weights of  a man

to that of the rat  (i.e., 83 kg/0.30 kg = 277. Ramsey and Andersen,  1984).

The 9% fat value for a 70 kg man  is an unrealistic reference physiological

value,  but represents a good fit  to the experimental data.  The  sum  of  the
                                   4-16

-------
tissue volumes represented 91% of  tie  total  body  weight  for  an 83 kg man.

     Droz (1985)  used the following tissue volumes  for his model

simulation:



                          Tissue	Pe r ce nt

                          Liver           2.4
                          Fat             16.0
                          VBG             10.0
                          Muscle group    52.0


These values are for a 70 kg man at rest.

     Andersen et al. (1987) reported the tissue volumes  for  a 70 kg man.

The volume percentages are:



                        Tissue	Percent

                        Liver                3.14
                        Fat                 23.1
                        Rapidly perfused     3.71
                        Slowly perfused     62.1


These percentages are in  close accord with data from the medical literature

(ICRP, 1975; Davis and Mapleson, 1981).  The 23.1% tissue fraction for fat

in humans is much closer  to  the ICRP recommendation of 19% than the 9% used

by Ramsey and Andersen  (1984).



4.3.  CARDIAC OUTPUT


      Cardiac output  is  defined as  the volume of  blood pumped by  each

ventricle of  the heart  per minute.  Cardiac output  can  be determined  by

multiplying the heart rate and the volume of blood ejected  by  each

ventricle during each beat  (Vander et  al.,  1975):
                                    4-17

-------
                Cardiac output  =  heart rate  z stroke vclone


The major determinant  of cardiac  output in all  species  is  the  oxygen demand

of the tissues.   During physical  activity,  cardiac  output  increases,  but  to

a smaller extent than  alveolar  ventilation (Astrand, 1983).  As  a  result,

the perfusion-ventilation ratio is decreased by exercise.   Thus,  the ratio

of cardiac output to alveolar ventilation decreases from a value of  1.3  at

rest to a value  of 0.4 to 0.5 with mild activity and a  value  of  0.4  with

strenuous activity (Astrand,  1983).



4.3.1.  Reference Values


     Several direct laboratory  measurements of  cardiac  output  in rats and

humans are available in the literature.  However, we were  only able  to

locate two direct measurements  of cardiac output in the mouse.  Based on

data available to us we are recommending the following  reference values  for

cardiac output (1/min):



                TABLE  4-4. REFERENCE CARDIAC OUTPUT VALUES
                   Mouse           Rat           Human
                (0.025 kg)      (0.25 kg)      (70.0 kg)

                0.017 1/min    0.083 1/min     6.2 1/min
4.3.2.  Documentation of Reference Values


     The reference cardiac output of 0.017 1/min for mice was calculated

using the power function suggested in section 5.1, using the reference

mouse body weight of 0.025 kg.  The reference cardiac output of 0.083  1/min
                                   4-18

-------
for the rat was also calculated using the power function suggested  in

section 5.1, using the reference rat body weight of  0.25 kg.
                                   Mice
                TABLE 4-5. ABSOLUTE CARDIAC OUTPUT OF MICE


       CO        Method             Ref.
       (1/min)

       0.0110    1                  Gjedde and Gjedde (1980)

       0.0118    2                  Wetterlin and Pettersson (1979)

       0.0160    -                  Blizard and Welty (1971)

       1. Indicator fractionation
       2. Soluble indicator



     Gjedde and Gjedde (1980) measured cardiac output in anesthetized,

female BALB/c mice weighing 0.025 kg.  Using the indicator  f ractiona tion

technique developed by Sapirstein  (1958), cardiac output was determined to

be 0.011 1/min.  Relative cardiac output was 0.440 1/min-kg.

     Wetterlin and Pettersson (1979) determined cardiac output in 35 male

and female Nlftl, anesthetized mice.  The mice weighed between 0.025 to

0.030 kg.  Cardiac output measured 0.0118 to 0.0028  1/min.   Relative

cardiac output determined from this  study was 0.435  1/min- kg.

     Blizard and Welty (1971) measured heart rates in 70 unane sthetized,

unrestrained mice with an average  body weight of 0.0225 kg.  Mean heart

rate 60 minutes after handling was 537.4 min"1-  Using an average stroke

volume of 0.03 ml (Wetterlin and Pettersson, 1979) for anesthetized mice,

we obtain an estimate of cardiac output  in mice  of 0.016 1/min, with a
                                   4-19

-------
relative cardiac output  of  0.711  1/min-kg.

     Table 4-6 summarizes  data  available  on the  relative  cardiac  output  of

mice.



                TABLE 4-6.  RELATIVE CARDIAC OUTPUT OF MICE
          	(1/min-kg)	

          £0      Range            Ref.

          0.440   -               Gjedde  and Gjedde (1980)

          0.435   0.333  -  0.537   Wetterlin and  Pettersson  (1979)

          0.711   -               Blizard and Welty (1971)
                                   Rats


     Measurements of absolute  cardiac output  of  unane sthetized rats are

listed in Table 4-7:
                                   4-20

-------
         TABLE 4-7.  ABSOLUTE  CARDIAC  OUTPUT OF UNANESTHETIZED RATS
CO
(1/min)
0.0730
0.0918
0.1010
0.1107
0.1167
0.1193
0.1340
Method
1
2
2
2
2
2
3
Ref.
Guy ton (1947)
Popov ic and Kent
Jansky and Hart
Tsuchiya et al .
Tsuchiya et al.
Tsuchiya et al.
Coleman (1974)


(1964)
(1968)
(1978)
(1978)
(1978)

         1. Respirograph
         2. Pick
         3. Dye-dilution
     Guy ton  (1947) used the oscilloscopic respirograph to determine  cardiac

output in 35 white rats with an average weight of 0.113  kg.   Cardiac output

was 0.0730 1/min.

     Popov ic and Kent (1964) conducted an extensive,  120-day study of

cardiac output in unane sthetized rats.  Using the Pick method,  'they

determined a cardiac output of 0.0918 1/min in rats weighing 0.319 kg.

Jansky and Hart  (1968) measured cardiac output in warm- and cold-acclimated

rats and reported a cardiac output of 0.1010 1/min.  Average weight of  the

rats was 0.355 kg.  Tsuchiya et al. (1978) measured cardiac output in three

strains of conscious rats.  Six male Wistar-Kyoto rats,  weighing 0.447  ±

0.020 kg,  gave an average cardiac output of 0.1107 1/min.  Eight male

hypertensive rats of the Akamoto-Aoki  strain, weighing 0.390 + 0.012 kg,

provided an average cardiac output of 0.1167 1/min.   Five male American

Wistar rats, weighing 0.473 ± 0.010 kg,  recorded a mean  cardiac output  of
                                   4-21

-------
0.1193 1/min.  All  experiments were  done  using  the  direct Pick principle.

Coleman (1974) measured cardiac  output  in  conscious Wistar rats weighing

0.512 ± 0.016  kg using a modified dye-dilution technique.  Cardiac output

was 0.134 1/min.

     Measurements  of  absolute cardiac output of  anesthetized rats are

listed in Table 4-8.



          TABLE 4-8.  ABSOLUTE CARDIAC OUTPUT OF ANESTHETIZED RATS
CO
(1/min)
0.027
0.040
0.047
0.047
0.063
0.070
0,079
0.093
0.095
0.097
0.100
0.120
Method
4
1
1 (a)
_
1. 6
5
3
3
3
3
3
2
Ref.
Walsh et al . (1976)
Walsh et al. (1976)
Blood et al. (1950)
Clarksoa (1956)
Chiu (1974)
Ac term a nn and Veress (1980)
Dytmdikova et «1 . (1981)
Dyradikova et al . (1981)
Lin et al. (1970)
Dyundikova et al . (1981)
Dyundikova et al . (1981)
Holt et al. (1968)
          1.  Pick (a)  a trial  cannula
          2.  Indicator-dilution
          3.  Theraodilution
          4o  Probe  method
          5.  Microsphere
          6.  Catheterization
                                   4-22

-------
     Walsh et al.  (1976) reported the cardiac output for 15 Spr ague-Daw ley




and 12 Wistar rats under basal resting conditions.   Using the direct Pick




procedure for determination of cardiac output,  Walsh et al. (1976)  compared




the results with simultaneous electromagnetic flowmeter techniques  (Probe




method).  Body weight ranged from 0.175 to 0.250 kg for both the Sprague-




Dawley and Wistar rats.  Pick cardiac output measurements in Spr ague-Daw ley




and Wistar rats under basal resting conditions were 0.041 1/min and 0.040




1/min, respectively.  Using the Probe method, Spr ague-Daw ley rats had a




0.029 1/min reading for cardiac output, while the Wistar rats measured




0.027 1/min.  Thus,  cardiac output using the Pick procedure was between 40




and 50% greater than with the Probe method.




     Blood et al.  (1950) determined cardiac output  in young adult,




anesthetized rats at normal and high altitudes.  The University of Denver




strain mice weighed between 0.180 to 0.200 kg.  The mean cardiac output for




52 rats at normal altitude was found to be 0.0465 ± 0.0156 1/min.  Claris on




(1956) reported cardiac output in an anesthetized,  0.18 kg rat as 0.047




1/min, with a range of 0.015 to 0.079 1/min.




     Chiu (1974) used a modified version of  the catheterization technique




popularized by Popovic and Kent (1964) and used again  by Jansky and Hart




(1968).  Chiu measured cardiac output in 25  male adult albino Wistar rats,




weighing 0.304 ± 0.005 kg, under  "conscious" conditions.   Cardiac  output




was 0.0625 ± 0.0028 1/min.  This measurement was obtained  when the  rats




regained consciousness following administration of  40  mg/kg  i.p. of  sodium




pentobarbi tal.  In compari son with other measurements  for  unane sthetized




rats,  it is clear the value reported by Chiu (1974)  is more  in line with




those  of anesthetized rats.  Chiu  (1974) suggested  cardiac output  should




not be affected by the residual effects of  light  anesthesia.   However,  the
                                   4-23

-------
amount of sodium pentobarbital  used for  the  rats is equal  to  the  amount




reported for anesthetized rats  in the study  by  Popovic and Kent  (1964).




Chiu  (1974)  did not specify the time lapse  between the administration of




the anesthetic and the regaining of consciousness.




     Lin et al„  (1970) measured cardiac  output  in anesthetized,  Sprague-




Dawley rats in an attempt to prove the reliability of  the  thermodilution




method for measuring cardiac output.  Cardiac output ranged from 0.066 to




0.124 1/min for  rats that ranged in weight  from 0.330 to 0.494 kg.   The




author notes that the thermodilution method results in higher cardiac




output values than those obtained by other  methods.




     Ackermann and Veress (1980) measured cardiac output in 24 male




Spr ague -Daw ley rats.  Body weight was 0.394  + 0.008 kg.  Cardiac output was




measured as 0.0701 1/min.  Dyundikova et al.  (1981) measured cardiac output




in male rats as  a function of age.  Rats weighing 0.345 kg (ages 5  to 9




months) gave a cardiac output reading of 0.100 1/min.   Ten to 16-month old




rats,  weighing 0.383 kg, gave a cardiac  output  reading of  0.097 1/min.




Seventeen to 23-month old rats, weighing 0.420  kg,  gave a  cardiac output




reading of 0.078 1/min.   Twenty-four to  29 month old rats, weighing 0.411




kg, gave a cardiac output reading of 0.093  1/min.  Thus, cardiac output




decreases with age, reaching a  minimum at 17 to 23 months. However, rats




older than 24 months approached the values found for rats between 17 to 23




months (Dyundikova et al. , 1981).  Holt  et al.  (1968) reported a cardiac




output measurement of 0.12 1/min  in a controlled experiment using  three




anesthetized rats weighing 0.49 kg.




     Relative cardiac output measurements of unane sthetized and




anesthetized rats are summarized  in Table 4-9  and Table 4-10.
                                   4-24

-------
TABLE 4-9 .  RELATIVE CARDIAC OUTPUT OF UNANESTHETTZED RATS
                       (1/min-kg)


CO      Range             Ref.

0.248   -                 Tsucniya et al.  (1978)

0.252   -                 Tsuchiya et al.  (1978)

0.265   0.252 - 0.278     Coleman (1974)

0.278   0.255 - 0.301     Malik et al.  (1976)

0.285   -                 Jansky and Hart (1968)

0.286   0.261 - 0.311     Popovic and Kent (1964)

0.299   -                 Tsnchiya et al.  (1978)

0.305   0.264 - 0.346     Malik et al.  (1976)

0.402   0.383 - 0.421     Muller and Mannesaann  (1981)

0.646    	Gnyton (1947)	
                           4-25

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TABLE 4-10. RELATIVE CARDIAC OUTPUT OF ANESTHETIZED RATS
                       (1/min-kg)
£0      Ranee               Ref.

0.204   0.186 - 0.222       Popovic and Kent (1964)

0.205   -                   Sapirstein (1958)

0.207   0.203 - 0.210       Chin (1974)

0.210   0.176 - 0.242       Bollard (1959)

0.228   0.213 - 0.243       Coleman (1974)

0.238   0.196 - 0.299       Richardson et al.  (1962)

0.245   -                   Holt et al.  (1968)

0.245   0.163 - 0.327       Blood et al. (1950)

0,249   0.240 - 0.258       Richardson et al.  (1962)

0.261   -                   Clarkson (1956)
                          4-26

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                                  Humans


     Measurements of absolute  cardiac  output  of  humans are  summarized in

Table 4-11.



              TABLE 4-11.   ABSOLUTE  CARDIAC OUTPUT OF HUMANS
CO
(1/min)
4.60
4.88
5.20
5.21
5.30
5.51
5.60
5.73
6.00
6.00
6.21
6.30
6.49
Method
1
6
4
2
4
5
-
6
3
5
6
-
2
Ref.
Shaw et al . (1985)
Starr and Shroeder (1940)
Frostell et al. (1983)
Astrand et al. (1964)
Reeves et al. (1961)
Cournand et al. (1945)
Guyton (1971)
Starr and Shroeder (1940)
Johnson and Miller (1968)
Stead et al. (1945)
Tanner (1949)
Cowles et al. (1971)
Brandf onbrener et al. (1955)
              1. Doppler
              2. Dye-dilution
              3. Breath-hoi ding
              4. Direct Pick
              5. Catheterization
              6. Ballistocardiograph
     Tanner (1949) reported cardiac output data for 50 healthy men measured

by the ball istocardiograph.  Mean cardiac output was 6.21 + 0.79 1/min.
                                   4-27

-------
Tanner also reported cardiac output  measurements  by  other  investigators  for




both males and females.   Ball istocardiograph measurements  ranged  from  4.88




to 6.21 1/min (Starr and Shroeder, 1940;  Tanner,  1949).  Catheterization




resulted in cardiac output  measurements of  5.51  1/min and  6.00  1/min (Stead




et al,, 1945; Cournand et al.,  1945).




     Guyton (1971) recommends a c&rdiac output  of 5.6 liters per  minute  for




a normal,  resting adult man.




     Astrand et al. (1964)  measured  cardiac output in 23  healthy  men and




women using the dye-dilution technique.   Cardiac  output  at rest ranged from




3.8 to 6.7 1/min.   Average  cardiac output was 5.21 1/min.




     Reeves et al. (1961) reported a measured value  of 5.3 1/min for a




resting 72 kg man.



     Johnson and Miller (1968)  reported a resting cardiac  output  of 6.0




1/min.  Shaw et al. (1985)  measured  cardiac output in 10 subjects using the




pulsed Doppler technique.  Resting cardiac  output was 4.6  ± 1.0 1/min




(range of 3.2 to 6.2 1/min).  These  measurements are in agreement with




those of Astrand et al.  (1964).




     Cowles et al. (1971) reported a cardiac output  of 6.3 1/min based on a




statistical analysis of 510 measurements by 45  investigators,




     Brandfonbrener et al.   (1955) measured cardiac output in 67 healthy




males, 19 to 86 years of age.  The mean cardiac output for a 23.6 year old




man was 6.49 1/min.




     The best summaries of data for  cardiac output in humans appear to be




Tanner (1949), Brandfonbrener et al. (1955). and Cowles et al. (1971).




These studies indicate that cardiac output decreases with age  but  is  about




6.2 to 6.5 1/min for men between the ages of 20  to 30 years  old.  Thus, we




are recommending a reference cardiac output  of 6.2  1/min.
                                   4-28

-------
4.3.3.  Values Used in Pharmacokinetic Models


     Table 4-12 summarizes cardiac output values that have  been used in

pharmacokine tic models.
                        TABLE 4-12. CARDIAC OUTPUT
                                  (1/min)
Source Mouse
Mapleson (1963)
Mun son and Bowers (1967)
Dedrick and Bischoff (1968)
Bischoff et al. (1971) 0.0044
Dedrick et al . (1972)
Dedrick et al. (1973) 0.0044
Harrison and Gibaldi (1977)
Lutz et al. (1977)
Fernandez et al. (1977)
Tuey and Matthews (1980) 0.0047
Ramsey and Andersen (1984) 0.015
Fiserova-Bergerova (1985)
Andersen et al . (1987) 0.039
Rat Human
6.48
5.00
6.48
0.03 4.04
4.04
4.04
0.0673
0.0244
6.5
-
0.0940 4.83
6.54
0.0850 5.80
                                    4-29

-------
4.4.  CARDIAC OUTPUT DISTRIBUTION


4.4.1.  Reference Values


     Cardiac output distribution depends  on the  oxygen demand  of  the

tissues which increases with physical  exercise  and decreases with

anesthesia (Astrand, 1983).   Blood flow rates,  like tissue  volumes,  can be

independently measured in the laboratory.   All  blood flow measurements

listed in this section are calculated in liters of blood per  hour and

presented as a percentage of total blood flow (cardiac output).
                TABLE 4-13 .  REFERENCE TISSUE HERFUSION RATES
                           (fraction of cardiac output)

Liver
Fat
VRG
MS
Mouse
0.25
0.09
0.51
0.15
Rat
0.25
0.09
0.51
0.15
Human
0.26
0.05
0.44
0.25
4.4.2.  Documentation of Reference Values



                                   Mice


     Wetterlin et al. (1977) reported measurement* of blood flow

distribution in anesthetized NMRI mice of both sexes.  Blood flow  to  the

VRG (brain,  heart, kidney, and viscera) accounted for 47.8% of total

cardiac output (brain,  2%; heart, 2.8%; kidney*, 14.6%; and the viscera,

30.2%).  Blood flow  to the liver measured 8.4 ± 0«8%  (arterial blood  flow
                                   4-30

-------
only).




     Quintan* et al.  (1979)  measured the distribution of  cardiac  output  in




BALB/c mice using the radioactive microsphere method.   The cardiac  output




distribution (%) was expressed in per gram of tissue.   We used the  organ




weights reported by Crispins (1975)  for this particular strain of mice.




When the organ weights were not available for the BALB/c  strain,  an average




of the reported values was used.   Blood flow to the liver was 21%.   The




total percentage of blood flow to the VKG was 54% (Brain, 13%;  Heart,  6%;




and kidneys, 35%).  No values were given for fat or the MB.






                                   Rats






     Sasaki and Wagner (1971) reported measurements of blood flow




distribution in unane sthetized rats.  Blood flow to the VUG (brain,  heart,




kidney, and viscera) accounted for 48.2% of the total cardiac output




(brain, 1.2 ± 0.31%;  heart,  2.9 + 0.43%; kidneys, 17.8 + 2.2%;  and  the




viscera, 26.3%).  Blood flow to the liver measured 6.7 ± 1.5% (arterial




blood flow only).  Malik et al. (1976) measured the fraction of blood flow




to the liver in 15 conscious Spr ague-Daw ley rats using the reference sample




method.  Blood flow was 20.26 + 1.9%.  Herd et al. (1968) reported an




approximate blood flow rate of 4  to 5% to adipose  tissue  in white,  male




Hoi tzman rats.




     Tsuchiya et al. (1978) determined regional  cardiac  output distribution




in three strains of unane sthetized  rats.  Rats  included  in this  study were




five male American Wistar rats (0.473 ± 0.010 kg); six male Wistar-Kyoto




rats (0.447 ± 0.020 kg); and eight  male hypertensive,  Okamoto-Aoki  rats




(0.390 + 0.012 kg). For  the  three strains of  rats, blood flow  to the VUG




accounted for 43.5% of total cardiac  output.  The  mean cardiac output
                                   4-31

-------
distributing into the liver  (includes  portal)  was  17%  for  the three strains




of rats.






                                  Humans






     Bradley et al.  (1945) reported a  human  liver  blood flow value of 26%.




Jones (1950) reported the  same  value  for  liver as  unpublished observations.




Leggett and Williams (1985)  designated 27% for liver blood flow.   Venous




blood accounted for  20% and  arterial  blood flow. 7.0%.




     Our fat blood flow distribution in humans was assigned a value of  5%




in accordance with the recommendation of  ICRP (1975).  Price et  al. (1960)




reported a body fat  blood  flow  fraction of 4.6%.   Cowles  et al.   (1971)




reported a body fat  blood  flow  fraction of 4.5%.




     The values for  the vessel-rich group are a compilation of measured




values suggested for the particular tissues  which  make up the  group.  The




tissues include the  brain,  heart,  kidney,  and viscera.  The blood flow




distribution of 13%  in the brain was supported by  measured values reported




by Eety and Schmidt  (1945).  Jone»  (1950),  and Chapman  and Mitchell (1965).




Brobeck (1979) recommended a slightly  higher value of  15%.  Blood flow




distribution to the  heart  in humans was measured at 4.0%  by Chapman and




Mitchell (1965) and  Brobeck  (1979).  Jones (1950)  suggested a  lower value




of 2.6%.   For the kidney,  a  blood  flow value of 23% was  reported by Smith




(1943) and Jones (1950).  Brobeck  (1979)  recommended  a lower value of 20%




and Chapman and Mitchell (1965) suggested 19%.  Edwards  (1975)  suggested an




inter species blood flow of 26%  of  cardiac output to the  kidney.   Brobeck




(1979) reported a measured value of 30% for the viscera  (includes liver).




Subtracting 26% of blood flow distribution for the liver results  in a blood




flow value of 4.0% for the viscera.
                                   4-32

-------
     Assam ing a blood flow distribution of  13% to the  brain, 4.0%  to  the

heart,  23% to the kidney and 4.0% to viscera,  results  in a  reference  value

of 44% for the VBG.   The range in this estimate  is 39  to 48%.

     Our definition of the muscle group includes the muscle and  skin.

Brobeck (1979) reported two values of 15 and 17% for  the muscle.   Chapman

and Mitchell (1965)  reported a higher value of 21%.   Blood  flow  to the  skin

was measured at 9.0% (Chapman and Mitchell, 1965) and  10.0% (Brobeck.

1979).   Cowles et al. (1971) reported 15.6% blood flow to the  muscle  and

skin of a 70 kg man.



4.4.3.   Values Used in Pharmacokinetic Models


                                   Mice


     Bischoff et al. (1971) reported blood flow rates for a 0.022  kg  mouse

as follows:



                             Tissue   Fraction

                             Liver      0.25
                             Kidney     0.18
                             Muscle     0.11


A total blood flow value of 0.0044 1/min was  used to  calculate the blood

flow percentages to the different compartments.  Blood  flow to the gut

(21%) was assumed to redistribute through  the liver.

     Dedrick et al.  (1973) provided  the following blood flow  rates for a

0.022 kg mouse:
                                   4-33

-------
                             Tissue    Fraction

                             Liver       0.41
                             Heart       0.06
                             Kidney      0.30
                             Lean       0.19
                             Marrow      0.04
Plasma values were converted to blood values  based on a  hematocrit  of 0.40,

and the blood flow rates for the liver,  kidney,  and gat  were  scaled in  pro-

portion to the data of Bischoff et al.  (1971).   They also  used a  total

blood flow value of 0.0044 1/min and assumed  the perfusion of  blood in  the

marrow tissue of mice was equal to that in the  rat.

     Tuey and Matthews (1980)  reported the following blood flow rates for  a

0.038 kg mouse :



                             Tissue   Fraction

                             Liver      0.41
                             Fat        0.01
                             Muscle     0.19
                             Skin       0.02


Blood flow to the muscle and liver were taken from the data of .Dedrick  et

al. (1973) and scaled per ml of tissue.  Blood flow values for the skin and

adipose tissue compartments were values scaled from a 0.250 kg rat (Lutz et

al., 1977) in proportion to the three-fourths power of the body weight

(Adolph,  1949).

     Ramsey and Andersen (1984) reported blood flow distribution in a 0.022

kg mouse  to the  following tissue compartments:
                                   4-34

-------
                        Tissue	Fraction

                        Liver               0.37
                        Fat                0.09
                        Richly  perfused     0.42
                        Muscle              0.12
Andersen et al.  (1987)  reported the  following blood flow fractions in a
0.038 kg mouse :
                        Tissue	Fraction

                        Liver                0.24
                        Fat                  0.05
                        Rapidly perfused     0.52
                        Slowly  perfused      0.19
                                   Rats


     Bischoff et al.  (1971) reported blood flow measurements  in a 0.200 kg

rat as:


  •
                             Tissue   Fraction

                             Liver      0.31
                             Kidney     0.24
                             Muscle     0.14


As with the nice, the measurements were based on a 0.40 heaatocrit  and

organ w«ifht-to-body weight relationships formulated by Adolph (1949).

     Harrison and Gibaldi  (1977) provided blood flow fractions for  a 0.360

kg rat as the following:
                                   4-35

-------
                        Tissue	Fraction

                        Liver                0.33
                        Kidney               0.27
                        Skeletal  muscle      0.16
                        Skin, fat, etc.        0.16
                        Heart                0.08
Total blood flow was 0.0534 1/min.   Blood flows  for  the  rat were  not  scaled

in this study due to differences in the  partition coefficients  of the

chemical between rats and humans.

     Lutz et al. (1977)  reported blood flow  distribution in a 0.250 kg  rat

to the following compartments:



                             Tissue   Fraction

                             Liver     0.19
                             Fat      0.05
                             Muscle   0.09
                             Skin     0.006



The fraction of blood distributing  into  the  skin is  a measurement of

effective blood flow.  The actual skin flow  rate used in the  model was

reduced by 10-fold in order to simulate  the  behavior of  the  chemical  under

investigation in the skin (Lutz et  al.,  1977).  The  blood values were in

close agreement with previously published data.

     Fiserova-Bergerova and Hughes  (1983) estimated the  following blood

flow rates for a 0.250 kg rat:
                                   4-36

-------
                             Tissue   Fraction

                             Liver     0.29
                             Fat       0.006
                             VBG       0.57
                             Muscle    0.07


Again, as with the fat volume of this model, the blood flow  to the  fat

appears to be underestimated.

     Ramsey and Andersen (1984) reported blood flow distribution in a 0.30

kg rat to the following tissue compartments:



                        Tissue	Fraction

                        Liver               0.37
                        Fat                 0.09
                        Richly perfused     0.42
                        Muscle              0.12



The blood flow to the liver, which represented 37% of cardiac output,

exceeded the more realistic value of 25%.  Ramsey and Andersen (1984)

surmised the need to increase the cardiac output was related to the

eztrahepatic metabolism of  the chemical under investigation.

     Andersen et al. (1987) reported blood flow rates in a 0.233 kg rat for

four tissue compartments:



                        Tissue	Fraction

                        Liver                0.24
                        Fat                  0.05
                        Rapidly perfused     0.52
                        Slowly perfused      0.19



These blood flow fractions were used for the mouse,  rat, and hman  species.
                                   4-37

-------
Ma pie son (1963)  reported  the blood flow distribution in a multi-

cdepartmental  model  for a 70 kg man that  included the following fractions:



                 Tissue	Fraction

                 Adrenals                         0.02
                 Kidneys                           0.19
                 Thyroid                           0.01
                 Gray  matter                       0.09
                 Heart                            0.04
                 Other small gland* and organs     0.01
                 Liver plus portal  system         0.25
                 White matter                      0.03
                 Red marrow                        0.02
                 Muscle                           0.09
                 Skin  a)  nutritive                 0.01
                      b)  shunt                     0.20
                 Non-fat  subcutaneous              0.01
                 Fat                              0.03



Total blood flow was 6.48 1/min.  Mapleson assumed that  the  blood flow  to

any tissue compartment is a fixed fraction of total  cardiac  output.

     Eger (1963) listed the following  perfusion rates for  a  70  kg man:



                            Tissue    Fraction
Liver
Fat
VHS
Muscle
VP6
0.26
0.05
0.49
0.18
0.02
Eger included liver in his vessel-rich compartment,  and the vessel-poor

compartment included bone and cartilage.

     Hanson and Bowers (1967) reported the distribution of blood flow in

human* as 75% to the VBD (included liver and brain), 19.6% for the muscle
                                   4-38

-------
group (includes bone),  and 5.4% to fat.   These  values  are  used  for

simulation of uptake during clinical  anesthesia.   If  liver  is designated as

a separate compartment,  these values  become:



                             Tissue   Fraction

                             Liver      0.26
                             Fat        0.05
                             VBG        0.49
                             MS         0.20



     Bischoff et al. (1971) reported the fraction of  blood flow in  a  70 kg

man as:



                             Tissue   Fraction

                             Liver      0.22
                             Kidney     0.19
                             Muscle     0.11



Total blood flow was 3.67  1/min.  Blood flow  to the gut (19%) was assumed

to redistribute through the liver.  As with the rodent data,  the blood

values are based on a hematocrit of 0.40 and organ weight-to-body weight

relationships.

     Dedrick et al. (1972) reported blood flow rates for a 70 kg man

essentially derived froa the work of Ma pie son  (1963).  The blood flow

fraction for each compartment was:
                                    4-39

-------
                            Tissue   Fraction
Liver
Kidney
Lean
Heart
Harrow
0.36
0.31
0.23
0.06
0.04
Dedrick et al.  (1973)  reported the  following blood flow rates for a 70 kg
man:
                            Tissue   Fraction
Liver
Heart
Kidney
Lean
Harrow
0.36
0.06
0.31
0.23
0.04
     Fiserova-Bergerova et al.  (1984)  used the  following blood distribution

fractions in her model simulation of  a 70  kg resting man:



                             Tissue    Fraction

                             Liver      0.23
                             Fat        0.05
                             VBG        0.45
                             Muscle      0.27



A separate compartment for the  lung was also  included  in  their model

simulation.

     Ramsey and Andersen (1984)  reported the  distribution of  blood flow  in

humans as equal to those values given for  the mouse  and rat:
                                   4-40

-------
                        Tissue	Fraction

                        Liver               0.37
                        Fat                 0.09
                        Richly perfused     0.42
                        Muscle              0.12
Andersen et al. (1987) reported the same blood flow fractions for a 70 kg

man as that of their 0.038 kg mouse and 0.233 kg rat:



                        Tissue	Fraction

                        Liver               0.24
                        Fat                 0.05
                        Richly perfused     0.52
                        Muscle              0.19



4.5.  VENTILATION


     Ventilation is a cyclic, dynamic process of circulation and exchange

of gases in the lungs that is basic to respiration.  Three primary concepts

used to quantify the ventilation  process are: minute volume, dead  space,

and alveolar ventilation.



Minute Volume


     The total volume of  air  exhaled per minute  is known  as  the  minute

volume or  total ventilation rate.



Dead Space


     Not all inhaled  air  is available  for  gas exchange  in the  alveolar

compartments.  The volume of  the  respiratory tract where  .no gas exchange
                                    4-41

-------
occurs is termed the dead space.   Anatomical  dead space is the internal

volume of the conducting airway from the nose and mouth down to the

alveoli.   The physiological dead space includes the anatomical dead space

and, in addition,  the conducting airways of the lower respiratory tract

where no significant exchange of oxygen and carbon dioxide between gas and

blood occurs.


Alveolar Ventilation


     The traditional definition of alveolar ventilation is the amount of

inspired gas which enters the alveoli per minute.  Thus, alveolar

ventilation is defined as:


    Alveolar ventilation = minute ventilation - frequency x dead space

                            •      •
                            Valv = VE - f x VD.


where VD is equal  to anatomical dead space.

     For the purposes of pharmacokinetic modeling, alveolar ventilation

should be defined as:

                            *      »
                            Vaiv - VE - f x VD.


where VD is equal  to physiological dead space.



4.5.1.  Reference Values


     Recommendations for reference respiration parameters are given in

Table 4-14.  The recommended minute volumes  are  calculated from  power
                                   4-42

-------
functions suggested in Section 5.1.  The alveolar ventilation rates are




calculated assuming alveolar ventilation is 67% of minute volumes for mice,




rats, and humans.  Physiological dead space is assumed to be  33% in mice,




rats, and humans.









               TABLE 4-14. REFERENCE RESPIRATION PARAMETERS







                                        	Species	
          Parameter                      Mouse     Rat      Human




          Body weight (kg)                 0.025    0.25    70.0




          Minute volume (1/min)            0.037    0.174    7.5




          Alveolar ventilation (1/min)     0.025    0.117    5.0




          Physiological dead space (%)    33.0     33.0     33.0




          Frequency (breaths/min)        163.0     85.0     15.0
4.5.2.  Documentation of Reference Values






     The reference minute volume of 0.037 1/min for mice was calculated




using the power function suggested in  section 5.1., using a reference mouse




body weight of 0.025 kg.  The reference minute volume of 0.174 1/min for




the rat was calculated using the power function suggested for rats in




section 5.1., using a reference rat body weight of 0.25 kg.  Alveolar




ventilation reference values for the mouse  and the rat were obtained by




taking 67% of the minute volumes for each species.  Physiological dead




space was calculated by taking  the remaining  33%  of minute volume for each




species.  The reference minute  volume  of 7.5  1/min for humans was taken




from ICRP (1975).
                                    4-43

-------
                                   Mice
Minute Volumes
     Absolute minute volumes of unanesthetized mice are summarized in Table
4-15.
        TABLE 4-15. ABSOLUTE MINUTE VOLUMES OF UNANESTBETIZED MICE
VE
(l/min)
0.024
0.028
0.049
0.052
Range

0.011
0.021
0.035
0.046

- 0.036
- 0.036
- 0.063
- 0.058
Method Ref.

1
2
3
3

Guy ton (1947)
Guy ton (1947)
Schlenker (1985)
Vinegar et al. (1979)
        1. Respirograph
        2. Head valve
        3. PI ethysmograph
     Guyton (1947) measured minute volume in 56 unane sthetized mice with an

average body weight of 0.02 kg.  Minute volume was 0.024 l/min with a range

of 0.011 to 0.036 l/min.  Schi enter (1985) measured minute volume  in

unane sthetized Swiss Webster mice with a mean body weight of 0.0304 +

0.0026 kg.   Minute volume was 0.0488 ± 0.014 l/min.

     Vinegar et al. (1979) measured minute volume  in male CD-I mice

weighing 0.027 ± 0.002 kg.  Using the body pie thysmograph, minute  volume

was measured at 0.052 l/min (range of 0.046 to 0.058 l/min).

     Absolute minute volumes of anesthetized mice  are summarized  in Table

4-16.
                                   4-44

-------
         TABLE 4-16. ABSOLUTE MINUTE VOLUMES OF ANESTHETIZED MICE
     VE            Range
     (1/min)

     0.021     0.009 - 0.046

     0.021     0.019 - 0.022

     0.052     0.038 - 0.066
Method
Ref.
2        Crosfill and Widdicombe (1961)

1        Vinegar et al.  (1979)

1        VonBucher (1949)
     1. PI ethysmograph
     2. Tracheal cannul a
     A minute volume of 0.021 1/min  (range of 0.009 to 0.046 1/min) for a

0.032 kg, anesthetized mouse was reported by Crosf ill and Widdicombe

(1961).

     Vinegar et al.  (1979) measured minute volume in anesthetized CD-I mice

and recorded a minute volume of 0.021  1/min  (range of 0.019  to 0.022

1/min).  The minute volume measurement  for the anesthetized nice

represented a 40% drop in minute volume  from the preanesthetic level.

     VonBucher (1949) measured minute  volume in 14 anesthetized mice.

Minute volume measured O.OS2 1/min  (range of 0.038 to 0.066  1/min).

     A summary of relative minute volumes of unanesthetized  and

anesthetized mice is presented in Table  4-17 and Table  4-18.
                                    4-45

-------
         TABLE 4-17.  RELATIVE MINUTE  VOLUMES  OF DNANESTHETIZED  MICE
                                 (1/min-kg)
•
VE Ranao Method
1.239 0.925 - 1.377 1
1.357 1.420 - 1.708 2
1.612 1.151 - 2.072 3
1.925 1.701 - 2.148 3
Ref.
Guy ton (1947)
Gay ton (1947)
Schlenker (1985)
Vinegar et al. (1979)
1. Respirograph
         2. Head valve
         3. PIethysmograph.
         TABLE 4-18.  RELATIVE MINUTE VOLUMES OF ANESTHETIZED MICE
                                (1/ain-kg)
      VE      Range           Method

      0.720   0.240 - 1.720   1

      0.777   0.704 - 0.815   2

      0.941   0.712 - 1.143   3

      2.26    1.652 - 2.869   2
Ref.

Crosfill and Widdicombe (1961)

Vinegar et al. (1979)

Guy ton (1947)

VonBueher (1949)
      1. Tracheal cannul a
      2. Pie thy tmo graph
      3. Tracheal valve
Dead Space


     The difficulty in obtaining accurate respiratory measurements in

animals smaller than rats precluded our obtaining sufficient data for mice.

Based on the interspecific consistency in anatomical dead space values, we

can conjecture physiological dead space values are also consistent between

species.  Therefore, we are recommending a physiological dead  space
                                   4-46

-------
reference value of 33% for mice.









Alveolar Ventilation






     We are recommending an alveolar ventilation reference  value  of  67% of




minute volume in mice based on corresponding data in rats and humans.








Frequency






     Measured respiratory frequencies of unanesthetized mice  are  summarized




in Table 4-19.









        TABLE 4-19. RESPIRATORY FREQUENCIES OF UNANESTHETIZED MICE
      (br/min)




      100




      163




      176




      186




      193




      213




      294*




      316*




      346*
                               Range
            Ref.
            Loosli et al.  (1943)




85 - 230    Guyton (1947)




157 - 195   Jaeger and Gearhart (1982)




143 - 226   Jaeger and Gearhart .(1982)




176 - 210   Jaeger and Gearhart (1982)




            Crossland et al.  (1977)




250 - 338   Jaeger and Gearhart (1982)




275 - 357   Jaeger and Gearhart (1982)




314 - 378   Jaeger and Gearhart (1982)
        Unrestrained animals
     Measured frequencies of anesthetized mice are  summarized in Table 4-
20.
                                   4-47

-------
         TABLE 4-20.  RESPIRATORY FREQUENCIES  OF ANESTHETIZED MICE
1 Range Ref.
(br/min)
109 97 - 123 Crosfill
154
168
198
210
-
and
Widdicombe (1961)
Drorbaugh (1960)
133 - 203 Morrison
- Morr i son
174 - 246 Morrison
80 - 200 Kl oilman
and
and
and
and
El sner
Eisner
Eisner
Radford
(1962)
(1962)
(1962)
(1964)
                                   Rats
Minute
     Tables 4-21 through 4-24 summarize the absolute  and relative minute




volumes of unanesthetized and anesthetized rats.




     Blame (1936)  measured minute volume in 23  unanesthetized rats weighing




0.111 kg.   Minute  volume was 0.228 1/min.   In 1943,  Blume and Zollner used




the head mask to determine minute volume in 14 unanesthetized rats.  Minute




volume measured 0.336 1/min for the 0.229 kg rats.




     Guyton (1947) determined the minute volume ia 35 white,  unanesthetized




rats (average body weight of 0.113 kg).  Minute volume was 0.073 1/min with




a range of 0.050 to 0.101 1/min.  Clarkson (1956) reported a minute volume




of 0.100 1/min (range of 0.075 to 0.130 1/min) for a rat  (body weight




unavailable).  Leong et al . (1964) determined the minute volume  for  four




groups of male, unane sthetized Wistar rats.  Minute volume rose  accordingly




with the increase  in body weight.  For 10  rats weighing  0.211 +  0.007 kg,
                                   4-48

-------
TABLE 4-21. ABSOLUTS MUTOTE VCLUIES OF UNANESTBETIZED RATS
VE        Method         Ref.
(1/nin)
0.057     4 (»)

0.073     1

0.087     1

0.089     4 (a)

0.094     1

0.100

0.113     1

0.115     3

0.147     4 (a)

0.150     4

0.153     7

0.154     4

0.161     4

0.162     1

0.169     4

0.174     4

0.186     4

0.189     3

0.189     4 (a)

0.194     4 (a)

0.215     2. 4

0.215     6

0.223     4

0.225     1

0.228     1

0.247     4

0.254     4

0.264     4

0.336     5
Bartlett and Tonney (1970)

Gnyton (1947)

Uoag «t al. (1964)

01 ton. aad D«ap»«y (1978)

Davis aad Morris (1953)

Clarkson (1956)

L«oag «t al. (1964)

Palxxk (1969)

Polianiki «t al. (1984)

Tboaas and Morgan (1969)

Natti* (1977)

M»nd«rly (1986)

Dorato »t al. (1983)

Lcoag «t al. (1964)

Laaa «t al. (1982)

Laadsy «t al. (1983)

Oorato •« al. (1983)

PalcMk (1969)

Polianski tt al. (1984)

Papp«ah«ia«r (1976)

Ito «t al.  (1976)

JUadcrlr «t al.  (1979)

Soil en tj aad H«ata  (1984)

L.ong «t al. (1964)

Bin* (1936)

TfceBas aad  Hortaa  (1969)

lUadcrly (1986)

H*nd»rl7 (1986)

Blnac aad Zollncr  (1943)
1. Reipiroirtph
2. Oas dilation, FSC
3. Boyle's Lav, FEC
4. Pl«thysaofrapli, (>) bazoB«trio
5. B*ad aask
6. Nonrebr»«tiing valr«s
7. B«ll<7*s puaoBOfraa
                     4-49

-------
TABLE 4-22. ABSCLDTE MINUTE VCLUJES OF ANESTHETIZED RATS
VE Method
(1/ain)
0.046 3
0.050 3
0,064 3
0.044 3
0.076 6
0.085 3
0.092 3
0.092 3
0.112 3
0.118 3
0.119 1
0.127 3. 5
0.131 3
0.146 1
0.149 3. 5
0.150 3
0.160 2
0.161 3
0.171 1
0.175 3
0.203 3
0.237 3. 4
0.38S 3
£•£.
Maudarly and Sifford (1978)
Mamderly and Sifford (1978)
EotMztson and Far hi (1965)
TnoBas and Morgan (1969)
Guy ton (1947)
Pal«c«k and Koch ova (1968)
Palaeck «t al. (1967)
Tnoaaa and Morgan (1969)
Paleo.k «t al. (1967)
Palaock and Chralora (1976)
Soneti (1960)
Cbralora »t al. (1974)
Pal»o»k (1969)
Schuti (1960)
Pal«
-------
TABLE 4-23. RELATIVE MINUTE VOLUMES  OF UNANESTHETIZED RATS
Relative VE
(l/«in-ki)
0.001
0.142
0.326
0.411
0.540
0.561
0.614
0.646
0.711
0.733
0.755
0.752
0.768
0.773
1.027
1.673
2.054
Method
6
3 (a)
3 (a)
5
3
7
8
1
5
2
1
4
1
3
1
1
1
Ifil-
Bluae tad Zollner (1943)
Bartlett and Tenney (1970)
Olson and Deapsey (1978)
Palecek (1969)
LaBM et al. (1982)
Mauderly et al. (1979)
Nattie (1977)
Guy ton (1947)
Palecek (1969)
Davis and Xosxit (1953)
Leon| et al. (1964)
Ito et al. (1976)
Leong et al. (1964)
Landxy et al. (1983)
L«ong et al. (1964)
Leong et al. (1964)
Blue (1936)
1. Respirograph
2. Respiroeieter
3. Pie thy 00 graph (a) baroaetric
4. Gas dilation, FRC
5. Boyle's Lav. FRC
6. Head »ask
7. Nonzebreathing valves
8. Bellows pneaogzaB
                          4-51

-------
TABLE 4-24. RELATIVE MINUTE VCLDJES OF ANESTHETIZED RATS
Relative VE
(I/Bin- kg)
0.170
0.201
0.239
0.393
0.415
0.434
0.435
0.457
0.472
0.595
0.600
0.627
0.635
0.637
0.691
0.691
0.707
0.730
0.886
1.630
Method
3
3
3
3
3
3
3
3
3
3
1
2
1
3. 5
6
3
3, 4
1
3
3
Rof.
Mauderly and Sifford (1978)
Robertson and Farhi (1965)
Mauderly and Sifford (1978)
Cavalova et al . (1974)
Palecek and Rochova (1968)
Palecek et al . (1967)
Johanson and Pierce (1971)
Palecek and Chvalova (1976)
Palecek et al. (1967)
Palecek (1969)
Sehuti (1960)
Crosf ill and f iddico.be (1961)
Schutz (1960)
Palecek (1969)
Gay ton (1947)
Diamond and 0' Donne 11 (1977)
Ito et al. (1976)
Schutz (1960)
Palecek (1969)
VonBttcher (1949)
1. Gas meter
2. Passive  inflation
3. Pie thy mo graph
4. Gas dilation, FRC
5. Boyle's Law, FRC
6. Tracheal valve
                        4-52

-------
minute volume was measured at 0.162 + 0.042 1/min.   The rats weighing 0.298




± 0.008 kg measured a minute volume of 0.225 ± 0.028 1/min.




     Landry et al.  (1983) measured minute volume in male Fischer 344 rats




with an average body weight of 0.225 kg.   At the optimal breathing capacity




for the rats, minute volume was measured at 0.174 + 0.039 1/min.








Dead Space






     Stahl (1967) predicted a dead space value of 0.0004 liters for a 0.250




kg rat.  Chvalova and Palecek (1977) measured respiratory dead space in




male Wistar rats as 0.00035 liters.  Ward et al. (1983) estimated dead




space  in Porton rats as 0.00034 liters.  A representative tidal volume for




a 0.250 kg rat is about 0.0015 liters  (Crosf ill and Widdicombe, 1961; Leong




et al., 1964; Dor a to et al., 1983).  Based on these data, anatomical dead




space  in rats is 23% of tidal volume.  This number is corroborated by Lai




et al. (1981) who estimated anatomical dead space in male Spr ague-Daw ley




rats to be 22% of tidal volume.




     The only study found estimating physiological dead  space  in rats was




Olson et al.  (1978), who estimated dead  space in male Spr ague-Daw ley rats




to be 37% of tidal volume.  However, our analysis shows  that anatomical




dead space values are consistent between species.  We conjecture that




physiological dead space  should also be  consistent between  species.




Therefore, we are recommending a physiological  dead space reference  value




of 33% of tidal volume for rats.
                                   4-53

-------
Alveolar Ventilation






     We are recommending an alveolar ventilation reference value of 67% of




minute volume for rats.









Frequency






     Respiratory frequencies of unane sthetized and anesthetized rats are




listed in Tables 4-25 and 4-26:
                                   4-54

-------
TABLE 4-25. RESPIRATORY FREQUENCIES OF UNANESTHETIZED RATS
I
(br/min)
60
82
84.9
85
86
91
94.5
102
109
110.2
116.1
119
119
121.4
130
131
134
136
139.3
139
142
142
152
152.5
Range
-
72 - 92
68.3 - 101.5
66 - 114
-
80 - 102
90.8 - 98.2
80 - 124
103 - 115
87.7 - 132.7
103.5 - 128.7
109 - 129
113 - 125
113.7 - 129.1
-
-
-
-
123.5 - 155.1
-
120 - 164
-
-
-
Ref.
McCuicJieon (1951)
Aggarwal et al. (1976)
Bartlett tad Tenney (1970)
Guy ton (1947)
Pappenheiaer (1976)
Palecek (1969)
Ito et »1. (1976)
Mauderly et al . (1979)
Olson and Dempsey (1978)
L«ong ot «1. (1964)
L«on| at tl. (1964)
Palecek (1969)
Nattio (1977)
L«ong at al. (1964)
ThoBas and Morgan (1969)
Maude rly (1986)
Blume and Zollner (1943)
Maude rly (1986)
Leong et al. (1964)
Thomas and Morgan (1969)
Lam et al. (1982)
Maud* rly (1986)
Bluve (1936)
Davis and Morris (1953)
                         4-55

-------
TABLE 4-26. RESPIRATORY FREQUENCIES OF ANESTHETIZED RATS


1          Range           Ref.
(br/min)

47         -               Mauderly and Sifford (1978)

48         -               Mauderly and Sifford (1978)

68         -               Thomas and Morgan (1969)

69         64.3 - 73.7      Robertson and Farhi (1965)

79         -               Thomas and Morgan (1969)

80         -               Drorbaugh (1960)

90         81 - 99         Vizek et al. (1975)

92.8       87.1 - 98.5     Chvalova et al.  (1974)

96         88.3 - 103.7    Palecek (1969)

97         84 - 126        Crosfill and ffiddicombe  (1961)

102        98.9 - 105.1    Ito et al.  (1976)

104        93 - 115        Palecek (1969)

108        79 - 137        Johanson and Pierce (1971)

108        98.7 - 117.3    Palecek and Chvalova  (1976)

110        -               Agostoni et al.  (1959)

112        103.3 - 120.7   Palecek (1969)

115        96 - 134        VonBncher  (1949)

115        94 - 136        Diamond and 0'Donne 11  (1977)
                           4-56

-------
                                  Human8
     Absolute minute volumes of humans are reported in Table  4-27
              TABLE 4-27. ABSOLUTE MINUTE VOLUMES OF HUMANS
•
VE Ranee
(1/min)
6.02
6.1
6.4
6.5 6.0 - 7.0
7.43 5.8 - 10.3
7.5
7.5
7.7
8.20* 5.38 - 9.14
8.73 4.9 - 12.2
9.0
Ref.
Tobin et al. (1983)
Frostell «t al . (1983)
Crosfill and Widdi combe (1961)
Stahl (1967)
Taylor (1941)
ICRP (1975)
Brobeck (1979)
Frostell et al. (1983)
Bergman (1967)
Guy ton (1947)
Mead (1960)
* Anesthetized
     Brobeck (1979) reported & minute volume of 7.5 1/min for a 70.0 kg




man.  ICRP (1975) also recommended a 7.5 1/min minute volume for Reference




Man.  Taylor (1941) measured minute volume in three male subjects and




reported a minute volume of 7.43 1/min  (range of 5.8 to 10.3) in a resting




state.  Guy ton  (1947) measured minute volume in a 68.5 kg man using the
                                   4-57

-------
oscilloscopic respirograph.   Minute  volume was 8.73  1/nin  (range of 4.9 to




12.2).  Crosfill and Widdicombe  (1961)  reported  a minute volume of 6.4  for




a 70 kg man.   Bergman (1967)  measured  a minute volume  range  of 5.38 to  9.14




1/min in anesthetized subjects breathing 23  to 30% oxygen.   Head  (1960)




reported an average minute  volume  measurement of 9.0 1/min for 27  subjects




at rest.  Stahl (1967)  reported  a  minute volume  range  of 6.0 to 7.0 1/min




for a 70 kg man.




     Relative minute volumes of  humans are  summarized  in Table 4-28.
              TABLE 4-28.  RELATIVE MINUTE  VOLUMES  OF HUMANS
      Relative VE   Range
Ref.
(1/min* kg)
0.0142
-
0.091
0.1071
0.1071
0.1265

0.0768 - 0.1306 Bergman (1967)
0.0857 - 0.1000 Stahl (1967)
Crosfill and Widdi combe (1961)
Brobeck (1979)
ICRP (1975)
0.0710 - 0.1768 Gnyton (1947)
Dead Space






     There are more extensive measurements of both anatomical and




physiological dead space in humans.   It is generally accepted that




anatomical dead space as measured by the Bohr equation at normal




ventilation is subject to over-estimation (Chakrabarti et al., 1986).  ICRP




(1975) recommended an anatomical dead space value of 0.160 liters and a




tidal volume of 0.75 liters for humans.  Based on these data, anatomical
                                   4-58

-------
dead space in humans is 21% of tidal volume.

     Nunn and Hill (1960) measured the ratio of physiological dead space to

tidal volume in 12 normal subjects and found the ratio to average 0.32.

Wasserman et al. (1967) found the ratio of physiological dead space to

tidal volume in 10 healthy males to average 0.33 under normal resting

conditions and 0.17 during exercise.  Hohsenifar et al.  (1985) state that

normal values for the physiological dead space to tidal  volume ratio are

0.33 at rest and 0.25 during mild exercise.  Based on these data, we are

recommending a physiological dead space reference value  for humans of 33%

of tidal volume.

     Table 4-29 lists anatomical dead space measurements for humans.



               TABLE 4-29. ANATOMICAL DEAD SPACE OF HUMANS
               VD                             Ref.
               (liters)

               0.130 + 16 « 40 Yr)   Tsunoda et al. (1972)

               0.136 ± 19 « 40 Yr)   Fowler (1949)

               0.147                  Tatsis et al. (1984)

               0.155 ± 24 (> 40 Yr)   Fowler (1949)

               0.156 ± 16 (> 40 Yr)   Tsunoda et al. (1972)

               0.160                  Svanb«rg  (1957)

               0.160                  ICRP  (1975)
                                   4-59

-------
Alveolar Ventilation






     Coaroe et al.  (1955)  recommended an alveolar ventilation rate  of  4.2




1/min in a normal,  adult male.   Head (1960)  reported an average  alveolar




ventilation rate of 5.2 1/min for 27 subjects at rest.   Fenn and Rahn




(1965) list an alveolar ventilation rate of  5.3  1/min.   Rackow et al.




(1965) recommended an alveolar  ventilation of 5.6 1/min based on two-thirds




of the expiratory minute volume of an anesthetized subject during the  first




five minutes of uptake.  Salzano et al.  (1984) reported alveolar




ventilation rates for divers at rest and during exercise.   The resting




values were taken while the divers inspired air containing 50% oxygen.  For




subject 1, resting alveolar ventilation was 7.0 ±0.9 1/min; subject 2,  6.8




±0.9 1/min; subject 3, 5.6 ±1.1 1/min; subject 4,  5.4 1/min; and subject




5, 6.6 ± 0.8 1/min.




     We are recommending an alveolar ventilation reference value in humans




of 5.0 1/min,  which is 67% of our.reference  minute volume.








Frequency






     Respiratory frequencies of humans are summarized in Table 4-30.
                                   4-60

-------
               TABLE 4-30. RESPIRATORY FREQUENCIES OF HUMANS


          1          Range         Ref.
          (br/min)

          10.3       8.1 - 12.7    Bergman (1967)

          11.7       10.1 - 13.1   Taylor (1941)

          14.2       10.5 - 19.3   Guyton (1947)

          14.8       -             Svanberg (1957)

          15         -             ICRP  (1975)

          15.6       -             Fowler (1948)

          16         -             Crosfill and Widdicombe (1961)

                     11-22       Stahl (1967)
4.5.3.  Values Used  in Pharaacokinetic Models


     Pharmacokine tic models use alveolar ventilation, rather than minute

volume, as an input  parameter.  Table 4-31 summarizes alveolar ventilation

values that have been used in pharmacokinetic models.
                                   4-61

-------
         TABLE 4-31. ALVEOLAR VENTILATION
                      (1/min)
Source                       Mouse    Rat     Human

Mapleson (1963)              -        -       4.0

Muason and Bowers (1967)      -        -       4«0

Ramsey and Andersen (1984)    0.0121   0.075   3.83

Fiserova-B. et al.  (1984)     -        -       5.80

Fiserova-Bergerova (1985)     -        -       5.3

Andersen et al. (1987)        0.039    0.085   5.80
                       4-62

-------
                                5. SCALING






     Many of the physiological parameters used in pharmacokine tic modeling




are directly correlated to the body weight of the particular organism.




These physiological parameters generally vary with the body weight




according to a power function expressed as:






                                y = a BW b






where y  is a physiological parameter of interest, and a and b are constants




(Fiserova-Bergerova and Hughes, 1983).  If the constant b equals one, the




physiological parameter y correlates directly with body weight.   If the




constant b equals two-thirds, the parameter y correlates with surface area.




This formula was taken from  the classical paper of Adolph (1949). which is




generally recognized as the  definitive source on the quantitative




relationship between body weight and the physiological parameters.




     The most desirable method of obtaining the physiological parameters




used in  a pharmacokine tic model is direct measurement.  When  such values




are not  available, necessary biological parameters for an untested species




can be obtained through scaling.  Scaling is defined as the orderly




variation of anatomic and physiologic properties with body weights (Adolph,




1949).   Animal scale-up is possible because both large and small  animals




are physiologically similar.  With only minor exceptions,  the mechanisms  of




absorption, renal excretion, distribution, and  storage are relatively




similar  in all species of animals, including humans  (Fiserova-Bergerova  and




Hughes,  1983).  The concept  of using body weight  scaling  in  pharmacokine tic




modeling was introduced by Dedrick (Dedrick, 1973; Dedrick and  Bischoff,




1980).   Many of the anatomical and physiological  parameters  can be
                                     5-1

-------
correlated among species as exponential  functions of  body  weight:
Property
Kidney weight
Brain weight
Heart weight
Lungs weight
Liver weight
Stomach and intestines weight
Blood weight
Constant
(a)
.0212
.081
.0066
.0124
.082
.112
.055
Exponent
(b)
0.85
0.70
0.98
0.99
0.87
0.94
0.99
            Adapted from Adolph (1949).








5.1.  CARDIAC OUTPUT






     Cardiac output is defined as the  volume of  blood pumped by  each




ventricle of the heart per minute.   Inter species scaling of  cardiac  output




is known to depend on the 0.75 power of  body weight  (Stahl.  1967;  White  et




al., 1968; Holt et al.,  1968;  Takezawa et  al.. 1980).  In fact,  there  is




considerable evidence that both cardiac  output and minute volume scale




across species with the  three-fourths  power  of body  weight (Kleiber, 1961;




Holt et al., 1968; Schmidt-Nielsen,  1970).   It can therefore be  assumed




that intraspecies extrapolations of  cardiac  output and minute volume should




also scale with the three-fourths power  of body  weight (Andersen et  al.,




1987).




     Correlations based  on empirical observations of inter species scaling




of cardiac output have found the following power functions:
                                    5-2

-------
                         TABLE 5-1.  CARDIAC OUTPUT
                                  (1/min)
Constant
(a)
0.102
0.162
0.166
0.187
Exponent
(b)
0.9988
0.76
0.78
0.81 .
Ref.
Patterson et
White et al.
Holt et al. (
Stahl (1967)

al. (1965)
(1968)
1968)

     It is known that these power functions slightly underpredict cardiac

output in small mammals (Guyton,  1947).

     Empirical measurements of cardiac output in mice,  rats,  and humans

follow.



5.1.1.  Mice
     Empirical measurements of cardiac output of mice are shown in Table
5-2.
                 TABLE 5-2. SCALING CARDIAC OUTPUT OF MICE
£0
(l/min)
0.0110*
0.0118*
0.0160
BW CO/BW 0.75 Ref.
(kg)
0.025 0.175
0.0272 0.176
0.0225 0.275
Gjedde et al. (1980)
Wetterlin et al . (1979)
Blizard et al. (1971)
* Anesthetized
                                    5-3

-------
Using the estimate  of  cardiac  output  in unane sthetized mice (Blizard and




Welty,  1971)  and assuming cardiac  output  is related to the 0.75 power of




body weight results in the following  power function, where cardiac output




is expressed in liters per minute  (1/min) and body weight  in kilograms




(kg):






                           CO  =  0.275 (BW) 0-75









5.1.2.  Rats






     Empirical measurements of cardiac output of  unane sthetized rats are




shown in Table 5-3.








         TABLE 5-3. SCALING CARDIAC OUTPUT OF UNANESTHETIZED RATS
co
(1/min)
0
0
0
0
0
0
0
.0730
.0918
.1010
.1107
.1167
.1193
.1340
BW
(kg)
0
0
0
0
0
0
0
.113
.319
.355
.447
.390
.473
.512
CO/BW 0.75 Ref.
0
0
0
0
0
0
0
.375
.216
.220
.203
.237
.209
.221
Guy ton (1947)
Popov ic and Kent
Jan sky and Hart
Tsuehiya
Tsuchiya
Tsuchiya
Coleman
et al.
et al.
et al.
(1974)
(1964)
(1968).
(1978)
(1978)
(1978)

     Assuming that cardiac output is related to the 0.75 power of body




weight and taking the geometric mean of the ratios of CO/BW 0.75 flom Table




5-3, results in the following power function for cardiac output in
                                    5-4

-------
unanesthetized rats,  where cardiac  output  is expressed in 1/min and body




weight in kg:






                           CO =  0.235  (BW)  °-75









Measurements of absolute cardiac output  of  anesthetized rats are listed in




Table 5-4.









          TABLE 5-4.  SCALING CARDIAC OUTPUT OF ANESTHETIZED RATS
£0
(l/min)
0.027
0.040
0.047
0.047
0.063
0.070
0.095
0.120
BW
(kg)
0.213
0.213
0.190
0.180
0.304
0.394
0.412
0.490
CO/BW 0.75
0.086
0.128
0.163
0.170
0.154
0.141
0.185
0.205
Ref.
Walsh et al. (1976)
Walsh et al. (1976)
Blood et al. (1950)
Clarkson (1956)
Chiu (1974)
Ac kern a nn et al. (1980)
Lin et al. (1970)
Holt et al. (1968)
     As SUB ing that cardiac output is related to the 0.75 power  of  body




weight and taking the geometric mean of the ratios of CO/BW °»?5 from Table




5-4. results in the following power function for cardiac output in




anesthetized rats, where cardiac output is expressed in 1/min and body




weight in kg:
                                    5-5

-------
                           CO = 0.150 (BW)  0-75









5.1.3.  Humans






     Measurements of absolute cardiac output  of humans are  summarized in




Table 5-5.








           TABLE 5-5.  SCALING ABSOLUTE CARDIAC OUTPUT OF HUMANS
CO
(1/min)
4.60
4.88
5.20
5.21
5.30
5.51
5.60
5.73
6.00
6.00
6.21
6.30
6.49
BW
(kg)
73
63
-
69
72
68
70
73
75
75
72
70
68
CO/BW 0.75 Ref.
0.184
0.218
-
0.218
0.214
0.233
0.231
0.229
0.235
0.235
0.251
0.260
0.274

Shaw et al. (1985)
Starr et al. (1940)
Frostell et al. (1983)
Astrand et al. (1964)
Reeves et al. (1961)
Cournand et al. (1945)
Guyton (1971)
Starr et al. (1940)
Johnson et al. (1968)
Stead et al. (1945)
Tanner (1949)
Cowles et al. (1971)
Brandf onbrener et al. (1955)
     The best summaries of data for cardiac output in humans appear  to be




Tanner (1949), Brandfonbrener et al. (1955). and Cowles et al.  (1971).




These studies indicate that cardiac output decreases with age but  is about
                                    5-6

-------
6.2 to 6.5 1/min for men between the ages of 20 to 30 years old.   Thus,  we

are recomaending a reference cardiac output of 6.2 1/min.



5.2.  MINUTE VENTILATION


     Ventilation is a cyclic process of circulation and exchange  of gases

in the lungs that is basic to respiration.  Total ventilation or  minute

volume is defined as the volume of air exhaled per minute.   Minute volume

has been correlated with the three-fourths power of body weight:
                         TABLE 5-6. MINUTE VOLU1E
                                   (1/min)
Constant
(a)
0.332
0.373
0.379
Exponent
(b)
0.74
0.75
0.80
Ref
Adolph (1949)
Guyton (1947)
Stahl (1967)
For mice, minute volume was shown to  scale, as was cardiac output, with a

mass exponent of 0.75.

     Empirical measurements of minute volume  in mice, rats, and humans

follow.



5.2.1.  Mice


     A summary of empirical studies of unane sthetized mice  for which  both

minute volume and body weight were available  is  provided  in Table 5-7.
                                     5-7

-------
         TABLE 5-7.  SCALING  MINUTE  VCLDHES  OF ONANESTHETIZED  MICE
VE
(l/min)
0.0245
0.0281
0.0489
0.0521
BW
(kg)
0.0198
0.0207
0.0304
0.0270
VE/BW 0.7J
0.464
0.515
0.672
0.782
* Ref.
Guyton (1947)
Guyton (1947)
Sehlenker (1985)
Vinegar et al. (1979)
     Assuming minute volume is related to the 0.75 power of body weight and
                                           *
taking the geometric mean of the ratios of VE/BW °-75 from Table 5-7

results in the following power function of minute volume in unane sthetized

mice, where minute volume is expressed in 1/min and body weight in kg:


                           VE = 0.595  (BW) 0-75



Measurements of minute volumes of anesthetized mice are listed in Table 5-

8.



          TABLE 5-8. SCALING MINUTE VOLUICS OF ANESTHETIZED MICE
VE
(l/min)
0
0
0
.021
.021
.052
BW
(kg)
0
0
0
.032
.027
.023
•
VE/BW 0-75 Ref.
0
0
0
.278
.315
.881
Crosf ill
and
Vinegar et al
VonBucher
Widdi combe
. (1979)
(1961)

(1949)
                                    5-8

-------
     As Sinn ing minute volume is related to the 0.75 power of body weight and
                                           •
taking the geometric mean of the ratios of VE/BW °-75 from Table 5-8,

results in the following power function of minute volume in anesthetized

mice, where minute volume is expressed in 1/min and body weight in kg:

                           •
                           VE - 0.426 (BW) 0-75



5.2.2.  Rats


     Table 5-9 presents data on minute volumes of unane sthetized rats.
                                     5-9

-------
TABLE 5-9.  SCALING MINUTE VCLU1ES OF UNANESTHETIZED RATS
VE        M      VE/BW
(l/ain)   (kj)
0.037     0.400   0.113

0.073     0.113   0.375

0.087     0.052   0.799

0.089     0.273   0.236

0.096     0.131   0.441

0.113     0.110   0.592

0.115     0.280   0.299

0.147     0.294   0.368

0.153     0.249   0.434

0.154     0.219   0.481

0.161     0.218   0.505

0.162     0.211   0.520

0.169     0.313   0.404

0.174     0.225   0.533

0.186     0.248   0.529

0.189     0.266   0.510

0.189     0.296   0.471

0.215     0.222   0.665

0.215     0.383   0.442

0.225     0.298   0.558

0.228     0.111   1.186

0.254     0.255   0.708

0.264     0.219   0.825

0.336     0.229   1.015
                               Bietlatt tad Tenney (197C)

                               Go?ton (1947)

                               L.OBI «t al. (1964)

                               01 ton and De»pt»y (1978)

                               Davis and Morris (1953)

                               L*ong «t al. (1964)

                               Ptlec«k (1969)

                               Polianski »t al. (1984)

                               Natti* (1977)

                               Maodasly (1986)

                               Dor«to «t al. (1983)

                               L.onj «t al. (1964)

                               LUB «t al. (1982)

                               Landry »t al. (1983)

                               Dor a to «t al. (1983)

                               Pal«o»k (1969)

                               Poliaaaki ct al. (1984)

                               Ito «t al.  (1976)

                               Jteiuterljr »t al. (1979)

                               Loonj it al. (1964)

                               Bloa. (1936)

                               Ma«d«rl7 (1986)

                               M»ud.rly (1986)

                               Bloa« and Zollacr  (1943)
                         5-10

-------
     Assuming that minute volume is related to the  0.75  power  of  body

weight and taking the geometric mean of the ratios  of  V£/BW 0.75  flom Table

5-9, results in the following power function for  minute  volume in

unane sthetized rats,  where minute volume is expressed  in 1/min and  body

weight in kg:

                            •
                            VE - 0.492 (BW)0.75



     Table 5-10 lists minute volumes of anesthetized rats.
                                    5-11

-------
          TABLE 5-10.  SCALING  MINUTE VCLDJES OF ANESTHETIZED RATS
VE
(I/Bin)
0,046
0.050
0.064
0.076
0.085
0.092
0.112
0.118
0.119
0.127
0.131
0.146
0.149
0.160
0.161
0.171
0.175
0.203
0.237
0.388
BW
(kg)
0.271
0.209
0.318
0.110
0.205
0.195
0.258
0.258
0.160
0.323
0.220
0.240
0.234
0.280
0.233
0.285
0.402
0.229
0.335
0.238
VE/BW °-75
0.123
0.162
0.151
0.398
0.279
0.314
0.309
0.326
0.470
0.296
0.408
0.426
0.443
0.416
0.480
0.438
0.347
0.613
0.538
1.139
Ref.
Mauderly et al. (1978)
Mauderly et al. (1978)
Robertson and Far hi (1965)
Guyton (1947)
Palecek and Rochova (1968)
Palecek et al. (1967)
Palecek et al. (1967)
Palecek et al. (1976)
Sehutz (1960)
Chvalova et al. (1974)
Palecek (1969)
Sehatz (1960)
Palecek (1969)
Crosf ill and Widdi combe (1961)
Diamond et al. (1977)
Sehutz (1960)
J chanson et al. (1971)
Palecek (1969)
Ito et al. (1976)
VonBucher (1949)
     Assuming that minute  volume  is related to the 0.75 power of body
                                                      •
weight and taking the geometric mean of  the ratios of  VE/BW 0.15 from Table
                                   5-12

-------
5-10, results in the following power function for minute  volume in


anesthetized rats,  where minute volume is expressed in 1/min and body


weight in kg:



                           VE = 0.359 (BW)  0.75




5.2.3.  Humans



     Minute volumes of humans are reported in Table 5-11.
VE
(1/min)

6.02

6.4


7.43

7.5

7.5

8.73

6.5
               TABLE 5-11. SCALING MINUTE VOLUMES OF HUMANS
                 Bf     VE/BW
                 (kg)
70


-


70


70
       0.265


       -


       0.310


       0.310
68.5   0.367

70     0.269
                                     Tobin et al .  (1983)

                                     Crosf ill and Widdicombe  (1961)


                                     Taylor (1941)


                                     ICRP (1975)


                                     Brobeck (1979)


                                     Guyton (1947)

                                     Stahl (1967)
                                   5-13

-------
                                APPENDIX A






     This appendix provides a limited review of the literature on partition




coefficients for use in pharmacokine tic modeling.  A partition coefficient




is an expression of the solubility of a chemical.  It is used to describe




the partitioning of a given chemical between air and blood, and blood and




tissues.  Partition coefficients can be determined in the laboratory using




a vial equilibration technique  (Sato and Nakaj ima, 1979).
                                     6-1

-------
    COEFFICIENT FOR MCE, fcAIS, AND HUKAKS AS REPORTED  IN  THE UIERATURE.



LIVES           FA1          nuSCU         KIDNEY         PLASM          LUN6
                 CMfniCAL
                                      SPECIES   Oil/ROOD
                                                                 OIL
                                                                               BLOOD
                                                                                                                                                                                 VESSEL RICH  VESSEL POOR   SERUIt    REF
Acetone
Acetone
Acrtont
Acttone
Acrtont
Acetylene
Acrtylrnt
Acrylonitrilt
Allylbtnirnt
Allylbtnirnt
Argon
tail Mr
Ininr
iMIIIt
InfMr
Irnme
Irnimr
Irnmr
Imint
Inline
Itnitnt
Imtme
ltni»t
tVoMk«oi(
-------

CHEHICAL
Cryofluorut
Cutcnt
Cuitnt
Cycloouui
Cyclohciut
Cycloproput
Cycloarount
Cyclopropui
Crclopropui
Cytlofroput
CfdoprofUi
Cytloyrouu
8icklarofemitM, i-
licklorobMim, •-
lickloroktflitm, o-
luklgrobnint, o-
licklorotthut 1,2-
Sicklorotthtnt, ,|-
licklorottkut, ,1-
licklorotthut, ,1-
ticklarotthut, ,1-
luklorottkut, ,1-
(7, tuklorottkut, ,1-
1 luklorottkut, ,2-
L'J ftcklorottkut, ,2-
licklorottkut, ,2-
lUklorottkut, ,2-
McklorMtkut, ,2-
fctHofMthylwe, cit-1,2-
hcklorotthylint, cit-1,2-
licklorMtkylMt, trui-1,2-
licklorottkylMt, trtat-1,2-
fccklorotthylwie, trui-1,2-
licklorofrofui, 1,2-
tickloroproptit, 1,2-
hitkyl itktr
liitkyl itlxr
Urtkyl ttkrr
dttkyl itlxr
tittkyl ttkir
kiftkyl tlkir
httkyl ttktr
lirthyl ttkir
Mitkfl tttoni
Mitkyl kttMii
li
-------

CHEKICAL
[nl lur *ne
f nf lur tat
tnf lurtnt
Eif lunnr
Enf lurtnt
Enflurtnt
Ethtnc
Elhtne
Ethine
Elhtnr
Ethyl chloridt
Ethyl chloridt
Ethyl chloridt
Ethyl forutt
Ethyl octint
Ethylbtnitnt
Ethylbtnitnt
Ethylbtnitnt
Ethyltnt
Ethyltfit
Fluorocirbon II
Fluorocirbon II
Fluorocirbon 113
fluorocirbon ||<
Fluorocirhoi 12
Fluoroctrbon 12
Fluroitit
Fluroitflt
fluroitnt
FluroiMt
fluroitflt
Hilothm
Hilothint
Htlothut
Htlothut
HtUthut
Htlothut
Hllothut
Htlothut
Htlothtne
Htftut, •-
H*tuont, 2-
HtitUuorotthut
Mnut, n-
luflurine
Iwfluruf
luflurint
IsoMurint
luf lurtnt
Krypton

SPECIES
huiin
huitn
huiin
huitn
huiin
rtt
huiu
huiin
huiu
MMII
huiu
huitn
huiu
huiu
huiu
huiin
huiu
hmu
huiu
huiu
huiu
huiu
huiu
huiu
hum
huiu
huiu
huun
huiu
huiu
huiu
huiu
huiin
huiu
huiu
huMi
huiu
huiu
huMJI
rjt
huiu
huiin
•OUIt
huiin
huiu
huiu
huiin
huiu
rit
•Dull

OIL/HLOOD OIL
...
...
...
...
...
...
...
...
...
II
—
—
24
...
...
37*1

133
—
...
27
...
32
5
— 3
—
—
...
...
—
...
...
...
—
...
—
—
—
—
...
4S2«/-20.0
4791
0.13
!44*/-4.f
—
—
...
...
...
0.50

BLOOD LIVES FST
2.0 2.3
1.7
l.» -- 70
1.3-2.2
l.t
2.o
0.10
O.OB
0.05
—
l.t
1.9

44.4
74.4
28.4
32.1
28.4»/-|.7
0.15 — O.tS
0.14 — 5.t
...
0.87
—
...
...
0.15
1.4 — 34
1.4
1.5
1.3
1.4
2.4 — 155
l.t
2.4
2.S
2.S — 40. 9
l.t-2.8
2.8
2.3-2.7
3.4
1. 9 1 /-O.I IO.BtM.0 J85»/-1J.«
Iff
— — —
0.80«/-O.OB 5.2«/-0.7 I04*/-l«.»
1.3
1.4 — 48
1.5
1.4-1.4
1.4
~~" --- ---
COdllNUfD.
rWSCLE KIDNEt PLASnA LUNG VESSEL RICH VESSEL POOH SCRUB HE
2
2
— — 1
2
2
2
2
2
2
3
1.2 4
2
2.3 — — — — 5
2
2
2
2
II
—
1.0 — — — 1.0 1.0
0.1
...
0.8
0.2
0.2
...
...
—
...
...
...
—
—
—
...
3.5 — — — 2.5 1.0
— — ... — ... —
—
—
...
12.5»/-0.4 8.t»/-l.2 — 2.5»/-0.2
— — — — — — —
— — — — — — —
5.0«/-0.» 3.0*M.I — l.0*/-0.07
2
1
2
2
2
3

-------
tn

CHEfllCAL
Hethinf
Htthoiyf lunnt
Httkoiyllurint
Httkoiyflurmt
Htthoiyflurmt
Hrthoiyflurinr
Htthoiyflurmt
Itctkoiyflurmt
Httkyl chloridt
Httkyl ckloridt
Httnyl chloridt
Httkyl ttkyl kttont
Httkyl ttkyl kttont
flttkyl liobutyl kttont
•ttkyl itofcityl kttont
Ittkyl i-kutyl kttont
Httkyl i-tutyl ktto»
llttkyl itityl kttont
Hrtkyl propyl kttont
Httkyl propyl kttont
HttkylcyclofHtiflt
HrtkyltM ckloridt
Httkyl Mt ckloridt
•ttkylnt ckloridt
flrtkyltut chloridt
•ttkylMt ckloridt
RttkylMi ckloridt
ftttkyltflt ckloridt
Hrtkyl tut ckloridt
Httkylhfiint, 3
•rtkylptfltttt, 2-
IWhylptMiM, 3-
IhMCkloraiitmtiit
Intrift*
totroMthint
fetrMt oiidt
fetroui otidt
lltroM and!
(btrmt oiidl
totrnt oiidt
fntiit, •-
h*yl ttktr
fr^ylitniMf
frt»ylktnit«t
Snot 1 unit
StyrtM
Styrtnt
Styrtnt
Styrtut
Styrtnt
	 	 - - •

SPECIES
huiin
huun
huiin
huiin
huiin
huiin
huiin
rit
hum
hunii
huiin
h«Mn
kuitn
huiin
hulUI
huiin
hum
hutn
hum
hiiin
hum
huiui
kutin
kuiin
ku»m
hum
huiin
kutn
rit
hum
hum
hutm
hum
•Mil
hum
hum
kutrn
hum
kum
MUM
huiu
h»«m
kum
hum
hum
hum
hum
hu»n
huiin
rit


OIL/BLOOD

---
---
...
...
...
—
...
...
---
...
...
1.3
10
...
...
13
34
4.2
...
...
...
...
...
...
...
...
...
...
—
...
...
...
...
...
...
—
...
...
—
...
—
208
—
...
...
105





OIL
...
...
---
---
...

...
...
...
8
—
243
—
...
924
...
—
...
...
424
202«/-1I.O
...
l52»/-5
—
75
—
152
—
...
3II»/-24.0
!03*/-7.3
HBt/-8.0
3743«/-»7
0.072
...
...
—
...
—
1.4
47«/-2.3
—
—
9775
53
...
...
4100
4114
jloj


CONTINUED.
BLOOD LIVER FAI IWSCLE HONE*
0.041
12.9
11-17
12.3
II --- 6/0
13.2
14-17
1
0.8
...
O.g — — —
202
202»/-IO
«•/-!!
to
—
!27»/-7
l««/-24
I50«/-I2
150
0. Bit/ -0.08 7.8«/-I.O I74«/-IO.O 5.0»/-0.8 4.7*/-|.l
7.0
9,7»/-0.4 — — —
7.0 — — —
—
5.3-4.4
9.7
9.4
12.8 — — —
I.]«H).I I0.4«/-l.l 277«/-24.0 I0.8W-0.7 7.3»/-2.l
0.4I»/-4.07 4.5«/-l.l 87»/-7.0 2.t«/-0.9 2.0»/-l.l
0.43«/-0.07 4.9«/-I.O 102»/-8.0 3.fl«/-0.8 2.5«/-I.O
30.8»/-l.9 — — — —
— ... — — —
42J
0.44 — — —
0.47 — 2.3 1.13
0.42
0.47 — 1.4 —
— — — — —
O.J8*/-O.Ofl 2.l»/'0.9 J9.4«/-2.4 0.7«/-0.4 0.4«/-0.3
8.8
47.0«/-2.4 — — —
47 ---
0.4
44
5I.9«/-2.0
IT 	 	 	 	
ii
j2 — — — —
40 2.7 50 1.0



PLASHA LUNb VESSEL RICH VESSEL POOR SERlffl REf
,
2
2
2
1
2
2
2
O.fl 4
1.7 — — — — 5
2
2
... ... ||
>.- - „
--- — 2
2
11
11
11
2
1.7»/-0.04 — — — 7
2
- I0
,.5 4
II — — |— — 5
2
2
_____ __ ___ 	 „_ 9
2
7
O.B*/-0.04 — — — 7
0.9»/-0.04 — — — 7
10
— — — j
— ... ... ... — . 2
2
1.04 1,0 — 4
2
— — — — — 1
3
0.5«/-0.03 — — — 7
— ... ... — 2
II
2
2
... ... ... 2
11
2
5.7 — — 8



-------

CHCIUCAl
Sulfur h»i«f luor lit
Sultur lunflLuridr
Svllur ktiifluwidt
Sulfur htiifluondt
UtrichlnroftkMt, 1, ,1,2-
!r(r«hlofo»llij«e, , ,1,2-
IttrichlorMthut, , ,1,2-
lttrichloro*tk«», , ,1,2-
Iftruhlorortkut, , ,2,2-
lttr«chlwMtkiM, , ,2,2-
lttruhloro*tk«nc, , ,2,2-
l«tr«klorottkwt, , ,2,2-
IrtrMhlorMtkylnt
!•( rtchl or o* Inline
IttrKhlorulkirlnt
ItUichlorottkfltM
TttrunlorottkjlMt
UtrMklvMtkyiiii
IkioMtkaivflirMC
lolMM
lolltM
III MM
Til MM
111 MM
III MM
1*1 MM
111 MM
1*1 MM
III MM
Til MM
IrickiorMtkiM, 1, , -
IrtctlorottkMC, 1, , -
IricklorMtktM, 1, , -
1ricUwM4kMe, 1, , -
IricklorMtkm, t, , -
TricklirwtlMM, 1, ,2-
TrictlirMtkuf, I, ,2-
TnrtlorMUm, 1, ,2-
IricUvMtlMM, 1, ,2-
IricklarMtkM*, I, ,2-
IricklffoctkfltM
tricklorMtbyliM
VicklirMtk|l«M
1ridtiWMtkvl«M
TncM«Mtk;l«M
trirtlvortkrlmi
Trirtlor«tth|lfi«
Iricklorortkylnt
ImklarMtkylMt

SCECIES
huun
tiuitn
kuitn
Mil It
lluiin
hunn
hunn
hunn
huiM
huitn
hMU
huiu
kuiu
kuaui
kuu
k«iM
kUMfl
kvttn
k»M
hum
kuw
kuin
kuin
kulM
ku«
kuMn
kUM
hUM
r«t
rit
hum
hum
kuin
hum
kuw
hutin
kuin
k.UM
kuiin
httAM
kUM
huu
kUM
kiun
nuiin
hunn
hunn
huain
hum

OIL/BLOOD Oil
...
—
—
0.30
4J06
...
43«4»M?5
I32I1«/-303
---
—
1110
13211
UI7
—
— —
4M
I1I7»/-I2»
—
7320
»4
1380
—
—
—
—
1471
...
...
...
...
...
354»/-20
...
I3»
334
380
...
2273«/-237
—
2273
—
7I«
—
...
...
718
718«/-8
---
"•"" ---
CONllNUED.
BLOOD LIVER FA! rlUSCLE MONEt PLASHA
0.00«
0.0064
0.0053
—
JO
72.6
J0.4*/-2.1
I2I.4«/-5.I
73
141
220
121
13.1
».l
II. t
32
I3.U/-I.I
».|
l£ 	 	 	
I3.4«/-I.7
13.4
7.3-13.1
U.j
l2f6.M»/-tl.32
14.;
13.6
14.24-84.5*
I4.34-J3.53
IS.2
t.4
l.«
3.3«/-0.4
1.4
7
j.j
56
44.2
38.4«/-4.4
44,2
31.4
t.»2«/-0.4J — 474.40«M4.08
».6
«.0 — 600
1.5
1.1
1.5
».5«/-0.6
».i
3.»-|0.2

LUN6 VESSEL RICH VESSEL POOR SERUn REF
2
2
2
3
2
78.2 4
10
10
2
2
5
2
2
2
2
5
JO
4
2
II
2
2
2
1
T.
2
13
13
2
2
3.4 4
10
2
— — — — ]
2
5
37.1 4
10
2
2
12
2
|
— 1
2
2
2
10
6
2

-------
CONTINUED.
CHEHICAL SPECIES OIL/BLOOD OIL BLOOD LIVER fAI HUSO.E KIDNEY PLASflA
If uMarotUyltne fit --- --- 26 --- --- 	 	
Irichlorotthyltnt rit — --- 25.B2«/-I.70 I.6H/-0.37 25.5HM.Ofl 0.43«/-0.01 l.35«/-0.48
Irichlorotthyltnt rit — — - 7.4
If ichlorotthyltnt, 1,1,2- huiin — 220 — --- — — — - 15
Ii ichlorotthyltnc, 1,1,2- huiin --- --- 9.5
lrHluoro-2-cklorocthint, 1,1,1- hu««i — 24 1.4
Itnon hiiMD — — 0.14 — 1.3
Itnon h««n — - 1.9 0.14
Itnon IOUM — 2.0 — — — — — —
lyltnt liuturt) kutM — 4030 42.1
lyltnt, •- hull* 146 — 24.4»/-0.9 — — —
lyltnt, i- kuitn — 3142 26.4
lyltnt, •- hiuu — — 24.50-30.98 — — ~
lyltflt, •- huitn — — — — 3404.47*/-I1B.7
lyltnt, i- (wain — — 21.0
lyltdt, •- kktu — — 27.12-30.47 — —
lyltftt, o- hiMD — 4340 31.1 — — — —
lyltnt, o- ku*M 140 — 3I.M/-2.J — — —
lyltnt, p- kuau — 3414 37.4 — — —
lyltnt, p- hum — — J4.2
lyltnt, p- hum 98 — 37.4*/-3.5 — — —
I
I. ftin|old, A. 1174. Antfth. Anilg. 55141:513-515.
2. Finfo»*-l»r(tra»i, V. 1983. Itodtlmi o< Inhilition Eipoiurt to Viporit Uptikt, Bittrikutim Hid ElUimtin 1:14-20. loci Riton fL:CK Prm.
3. Killer, K. *.; Piton, H. 1. li.j Stitk E. 1.) Suth, R. A. 1972. Anntkmol. 34(41:331-331.
4. Nor«M, A.; »Uck. A.; Itkhir, 0. R. 1170. Ann. Occuf. Hyj. 13:211-233.
5. tofjtfl, A.; 81 act, A.) Itilth, H.| Idckir, D. R. 1172. lit. i. Appl. I^Ut. Itat. 23i283-211.
4. Pipptr, E. H.i Mill, R. J., (di. 1143. Uptik* ind Ciitribitiw e< AtMtkttic Agjnts, pp. 71, 77. MM forki IkSrM-Hili leek Co.
7. Ptffetllix, I.; lru««ont, S.j Ctrttti, D.; hVintllt, t. 1183. Ir. I. Ind. ted. 42il42-167.
8. RiiitY, i. C.i Andtrioi, H. t. 1184. Toiicol. Appl. Pkiri. 73:139-173.
9. Site, .; Fujioirt, 1,; ktkijiu, T. 1174. jpn. 1. Ud. H*iith 16:30-31.
16. Site, .; lUkijiu, T. 1171. Arck. En.irofl. Htilth 34:41-73.
11 S«to | lUktjiM T 1171. Ir. J. Ind. tod. Ui231-234.
12. Sato, .; Hikiiiu, 1.; Fu|iwr«, ?.; Duriyui, N. 1177. Ir. J. Ind. tod. 34:36-41.
13. Site, ., ti it. 1172. Jpn. i. Ud. Htilth 14:3-1.

LUNG VESSEL RICH VESSEL POOR SERUN Rff
2
I.03«/-O.I7 — — — 12
2
5
5.1 4
2
2
2
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2
13
9
2
13
2
11
2
2
11



-------
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