Table Of ContentMETHODS IN
INHALATION
TOXICOLOGY
Edited by
Robert F. Phalen, Ph.D.
Boca Raton London New York
CRC Press is an imprint of the
Taylor & Francis Group, an informa business
CRC Press
METHODS IN THELIFESCIENCES
GeraldD. Fasman -
Advisory Editor
Brandeis University
Series Overview
MethodsinBiochemistry
JohnHershey
Departmentof BiologicalChemistry
Universityof California
CellularandMolecularNeuropharmacology
Joan M. Lakoski
Department of Pharmacology
Penn State University
Research MethodsforInbredLaboratoryMice
JohnP.Sundberg
The Jackson Laboratory
BarHarbor, Maine
MethodsinNeuroscience JosephM. Corless
SidneyA. Simon DepartmentofCell Biology,
Department ofNeurobiology Neurobiology and Ophthalmology
Duke University Duke University
MethodsinPharmacology
JohnH. McNeill
Professor andDean
FacultyofPharmaceutical Science
The University ofBritish Columbia
MethodsinSignal Transduction
JosephEichbergyJr.
Department of Biochemical and Biophysical Sciences
Universityof Houston
Methodsin Toxicology
EdwardJ.Massaro
Senior Research Scientist
National Health and EnvironmentalEffects Research Laboratory
Research Triangle Park, North Carolina
METHODS IN TOXICOLOGY
EdwardJ.Massaro -Advisory Editor
Senior ResearchScientist
NationalHealthandEnvironmental EffectsResearch Laboratory
Reasearch Triangle Park,North Carolina
The CRC PressMethods in Toxicology Series provides the reader with a
step-by-step approach to each ofthe up-to-date methods and techniques
which arepresentedinaclear andconciseformat. Topics coveringallaspects
ofthe methodsthat impacton toxicologyarebeing reviewed for publication.
Published Titles
Methods inRenal Toxicology,Rudolfs K. Zalups and Lawrence H. Lash
Molecularand CellularMethods in Developmental Toxicology,
George P.Daston
Methods in Inhalation Toxicology,Robert F.Phalen
Forthcoming Titles
PCR Application toMolecular Toxicology, JackVanden Heuvel
CRCPress
Taylor & Francis Group
6000 B roken Sound Parkway N W, Suite 300
Boca R aton, FL 33487-2742
First i ssued in h ardback 2 019
© 1 997 by T aylor & Francis Group, LLC
CRC Press is an imprint ofTaylor & Francis Group, an Infonna b usiness
No claim to original U .S. Government works
ISBN-13: 978-0-8493-3343-9 (Pbk)
ISBN-13: 978-1-138-47368-3 (hbk)
This book contains information obtained from authentic and highly regarded sources. Reasonable
efforts h ave b een m ade t o p ublish r eliable d ata a nd i nformation, b ut t he a uthor a nd p ublisher c annot
assume r esponsibility for t he validity of all materials o r the consequences of their use. T he authors and
publishers h ave a ttempted t o trace the copyright holders o f all material reproduced i n this publication
and apologize to copyright holders if permission t o publish in this form has n ot been obtained. I f any
copyright m aterial h as n ot b een a cknowledged please w rite a nd l et us k now so w e may rectify i n a ny
future r eprint.
Except as permitted u nder U .S. Copyright Law, no part o f t his book may be reprinted, reproduced,
transmitted, or u tilized in any form by any electronic, mechanical, or o ther m eans, now known or
hereafter i nvented, i ncluding p hotocopying, m icrofilming, a nd r ecording, o r i n a ny i nformation s tor
age o r r etrieval s ystem, w ithout w ritten p ermission f rom t he p ublishers.
For permission t o p hotocopy o r u se m aterial e lectronically f rom this w ork, please a ccess www.copy
right.com (http://www.copyright.coml) or c ontact t he C opyright C learance C enter, Inc. (CCC), 222
Rosewood Drive, Danvers, MA 0 1923, 978-750-8400. CCC is a not-for-profit organization that p ro
vides licenses and registration for a v ariety of users. F or organizations t hat have b een granted a p hoto
copy license b y t he CCC, a s eparate s ystem o f payment has b een arranged.
Trademark Notice: P roduct o r corporate names may be t rademarks o r registered trademarks, and are
used o nly f or identification a nd e xplanation w ithout i ntent t o i nfringe.
Visit the Taylor &. Frands Web site at
http://www.tayIorandfrancis.com
and the eRC Press Web site at
http://www.crcpress.com
Cover design: Dawn Boyd
Library of C ongress Card Number 9 6-25496
Library o f Congress Cataloging-in-Publication Data
Methods in inhalation toxicology I edited by Rober! F. Phalen.
p. cm. --(CRC Press methods in toxicology)
Includes bibliographical references and index.
ISBN 0-8493-3343-1
1. Pulmonary toxicology--Animal models. 2. Air--Pollution
-Toxicity testing. I. P halen, Rober! F., 1940- . H. Series.
[DNLM: I. R espiratory System--drug effects. 2. Toxicology
-methods. 3. Air P olIutants--toxicity. WF 100 M592 1996]
RCnO.M48 1996
615.9'02--dc20
DNLMlDLC
for Library of C ongress 96-25496
CIP
Abstract
Today, and in the foreseeable future, an appropriate understanding of the
biomedical effects of potentially significant natural and anthropogenic air
contaminants will require conducting inhalation exposure studies employing
whole living subjects. Conducting such studies is aspecializedresearch activ
ity that draws upon skills from avariety of scientific disciplines. How knowl
edge from these disciplines is integrated into the design and conduct of
inhalation studies is the subject of this book, which was written by an expe
rienced, active team of research scientists. Each of the vital aspects germane
to the conduct of such studies is discussed in detail, including the selection
of animal subjects and the related quality-control and ethical considerations,
airpurification methodology, exposure atmosphere generation and character
ization, inhalationexposure systems, andreal-time andpost-exposure biolog
ical assessments. The book, which is targeted to students in toxicology
programs andscientists contemplatingperforminginhalation studies,includes
comprehensive lists andaddresses of suppliers ofuseful equipmentandmate
rials relevant to each of the major aspects involved with the performance of
valid, reproducible inhalation toxicology studies.
Preface
Welive and breathe immersedina seaofair.Inaddition to themajor gaseous
components, oxygen and nitrogen, countless additional gases, vapors, and
particles are always present in the atmosphere and are available to deposit in
the respiratory system. Some ofthese contaminants arenatural, andsome are
anthropogenic. Of the anthropogenic substances, some arebeneficial (such as
aerosolized medications), and some are harmless or harmful depending upon
their concentrations. Ifwe are to maximize human wellbeing, we must study
andunderstand thebiomedicaleffects ofeachofthe potentially significantair
contaminants.Today, andinthe foreseeable future, this study andunderstand
ing will require conducting inhalation studies using whole living subjects. In
some studies, the subjects will be human volunteers, but in most studies
laboratory animals will be involved. Conducting such studies is a specialized
research activity that draws upon skills taken from numerous scientific disci
plines. These disciplines include chemistry, physics, engineering, biology,
physiology, microbiology, medicine, pathology, and others as well. How
knowledge from these areas is integrated into the design and conduct of
inhalation studies is the subject of this volume.
Thechapters in thisbookwerewritten byanactiveteamofscientistswho
have worked together in a modem inhalation toxicology program at the Air
Pollution Health Effects Laboratory at the University of California, Irvine.
Performing such studies has been amajor activity ofthis team for the past 10
years. In addition, these authors have trained students in research methods,
and they have established and taught several formal courses related to the
topics in this book. In short, each author is writing about his current daily
activities and within the scope of his formal training and experience. It is
hoped that this volume will provide a solid foundation for the readers — a
foundation that will support further specialized learning and will lead to
success in conducting theirown inhalation studies.
In addition to the authors, several people have contributed significantly
to the material herein. The chapters were reviewed and improved by the
technical staffof the Air Pollution Health Effects Laboratory. Special thanks
are due to Richard C. Mannix, M.S.; Charles Bufalino, M.S.; LauraA. Hoff
man, B.A.; and Michael J. Oldham, M.S. Additional editing, checking, for
matting, and word-processing were performed by Marie C. Tonini. Finally,
this bookis dedicatedwith gratitude andrespectto the research subjects; they
have been part of the team as well.
RobertF.Phalen, PhD.
Irvine, CA
The Editor
Robert Franklynn Phalen, Ph.D., directs the Air Pollution Health Effects
Laboratory at the University of California, Irvine (UCI). He also holds two
academic appointments in the College of Medicine at UCI: Professor in the
Department ofCommunity andEnvironmental Medicine andProfessorin the
Center for Occupational and Environmental Health (in the Department of
Medicine). He is amember of the faculty for the Graduate Program in Envi
ronmental Toxicology at UCI, and he teaches medical students, medical res
idents, graduate students, and undergraduate students in courses at several
universities including UCI, UCLA, and USC.
Dr. Phalen graduated in 1964 from San Diego State University with a
B.S. in physics (and a mathematics minor) and from the same institution in
1966 with an M.S. degree in physics with a specialty in nuclear physics. In
1971, he obtained a Ph.D. in biophysics, with specialization in inhalation
toxicology, from the University of Rochester (Rochester, NY). His postdoc-
torial research inaerosol physicsand inhaledparticledepositionmodelingwas
conductedatthe Inhalation ToxicologyResearch InstituteinAlburquerque, NM.
In 1972, Dr. Phalenjoined the College of Medicine at UCI to establish
the Air Pollution Health Effects Laboratory, which has since conducted over
100 studies relating to the toxicology of air pollutants. These studies have
been supported by avariety of federal agencies (including the U.S. EPA; the
National Heart, Lung and Blood Institute; the National Institute of Environ
mental Health Sciences; andtheCenters forDisease Control andPrevention),
California state agencies (California Air Resources Board,the Department of
Health Services, and others), and numerous private agencies (including the
Electric PowerResearch Institute, the Health Effects Institute, the Charles C.
Stocking Trust Fund, Southern California Edison Co., the Nickel Producers
Environmental Research Association, and others). His research has covered
several topics, including respiratory tract defenses, comparative respiratory
tract anatomy, basic aerosol science, inhaled aerosol deposition models, par
ticle and gas toxicity, andthe inhalability of large particles.
