Table Of ContentACS SYMPOSIUM SERIES 646
Environmental
Immunochemical Methods
Perspectives and Applications
g 01
acs.or6.fw0
bs.64 Jeanette M. Van Emon, EDITOR
u0
p://p996- U.S. Environmental Protection Agency
8, 2012 | htt0.1021/bk-1 Clare L. Gerlach, EDITOR
October 6 | doi: 1 Lockheed Martin Envrionmental Systems
32 on 3, 199 Jeffre C. Johnson, EDITOR
6.152.ober 2 U.S. Environmental Protection Agency
7.6Oct
ed by 21n Date:
do
oaati Developed from a symposium sponsored
ownlublic by the U.S. Environmental Protection Agency
DP
at the National Immunochemistry Summit IV,
Las Vegas, Nevada, August 2-3, 1995
American Chemical Society, Washington, DC
In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
QP 519.9 .I42E56 1996 Copy 1
Environmental immunochemical
methods
Library of Congress Cataloging-in-Publication Data
Environmental immunochemical methods: perspectives and applications /
Jeanette M. Van Emon, Clare L. Gerlach, Jeffre C. Johnson, editors.
p. cm.—(ACS symposium series, ISSN 0097-6156; 646)
"Developed from a symposium sponsored by the U.S. Environmental
Protection Agency at the National Immunochemistry Summit IV, Las
Vegas, Nevada, August 2-3, 1995."
g 01
acs.or6.fw0 Includes bibliographical references and indexes.
bs.64 ISBN 0-8412-3454-X
u0
8, 2012 | http://p0.1021/bk-1996- 3PII.rI I1o.E .t. JenVcIovmtahiirnonmo nsnEuo mAnnmo,geo aJnensnets,afca fJlyry e—e.m a CVnoCne..o,ti ttnN1oeg9ra rM5itne7isgo.s,- —ne 1sa.9Tl 5 .Ie 62mcI-h.V mnP.iuq oU.nul loenuIl—Iotiat.g enCyGdt so SeS—nrutglamaArtceemnhssas,.ie l tCysE s(.l4 inastr—vheir: CoL n.1o, m9n91ge59rn:e4t s8asLe-l as.s
32 on October 3, 1996 | doi: 1 V6Q2eP8g5.5a1—s9,. 9NdIc4e2v20.E) 56V I. 1S9e9r6ie s. 96-2817C9IP
6.152.ober 2
7.6Oct This book is printed on acid-free, recycled paper.
ed by 21n Date: Copyright © 1996
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oaati American Chemical Society
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PRINTED IN THE UNITED STATES OF AMERICA
In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
Advisory Board
ACS Symposium Series
Robert J. Alaimo Cynthia A. Maryanoff
Procter & Gamble Pharmaceuticals R. W. Johnson Pharmaceutical
Research Institute
Mark Arnold
University of Iowa Roger A. Minear
University of Illinois
David Baker at Urbana-Champaign
g 01 University of Tennessee
acs.or6.fw0 Arindam Bose AOTm&kTa Braemll L aNbaolraamtoraisesu
ubs.064 Pfizer Central Research
8, 2012 | http://p0.1021/bk-1996- MNRaoavbraeyl rR Ete .sF eCa. raBcshrta eLdllayibo,o nrJa rt.o ry NUVGonierniovtchererg sCnietaty r W Poolefin. c MaRo riSocahtbariegtoear tnUs n iversity
October 6 | doi: 1 MChaemrgEadriett C Aom. Cpaanvya naugh JUonhivner sRity. Sohf aIlplinleoyis
32 on 3, 199 National Science Foundation at Urbana-Champaign
7.66.152.October 2 AUnritvheursrit yB o. fE Wlliissco nsin at Madison DCoonucugrlaresn tA T. eScmhniotlhog ies Corporation
ed by 21n Date: GUnuivnedrsait yI .o fG Keaonrsags DL.u PSoonmt asundaram
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Lawrence P. Klemann William C. Walker
Nabisco Foods Group DuPont
Douglas R. Lloyd Peter Willett
The University of Texas at Austin University of Sheffield (England)
In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
Foreword
IHE ACS SYMPOSIUM SERIES was first published in 1974 to
provide a mechanism for publishing symposia quickly in book
form. The purpose of this series is to publish comprehensive
books developed from symposia, which are usually "snapshots
in time" of the current research being done on a topic, plus
some review material on the topic. For this reason, it is neces
g 01 sary that the papers be published as quickly as possible.
bs.acs.or646.fw0 propoBseefdo rtea bal es yomfp coosinutmen-btsa siesd r bevoioekw eisd pfuotr uanpdperro pcroinattreancets,s t htoe
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8, 2012 | htt0.1021/bk-1 Tproahupinse rda noisou ntp ytemhereo- ursescv oirpeewev eioedwf t phpreri oovrco eltsuosm fieisn. asIuln pa eacrdcvedipsiettaidonn cb,e ya odthrra erf eto jroegfc ateinoainczh.
October 6 | doi: 1 Terh^e ) aouft hthores sythmepno srieuvmise, wthheoi rb epcaopmeres thacec oerdditionrg( st)o otfh teh er ebcooomk .
32 on 3, 199 mcaemnedraat-iroenasd yo cf opbyo,t ha ntdh esu brmeviiet wtheres finanald ptahpee rse dtioto rths,e perdeiptoarrse,
ed by 217.66.152.n Date: October 2 wvtiioehnwos Acpohsafe pcpaekr re rstvuh ialaoerut,e sa olyilnnl lpcnylueu cbodelrseiissdgahi reniynda lprt heraveepis eseviraoosrnl cuashmr he eap nsva.oep t e Vbraesece rcnabenp amttdieam dod. re ri.eg pinraodl urec
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In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
Preface
IHE ENVIRONMENTAL ANALYTICAL COMMUNITY has responded with
enthusiasm to the need for reliable, inexpensive, field-transportable
methods for the characterization of hazardous waste sites by adapting
antibody-based technologies. Immunochemical methodology, with a
strong record in clinical applications, was a natural for the analytical chal
lenges faced by environmental scientists. There is a need for economical
org 001 methods that can detect trace levels of hazardous compounds in complex
s.pr
ac6. media such as soil, sludge, oil, and food products. Immunochemical
http://pubs.k-1996-064 tminose ttarh uvomadrsei enattyrae toi ocf namp. aabtIrlniec eosaf,d aadncithdioi ernve iqntugoi rveeth rliyetst lleoe w ifne da ttehuteerc ewtsi,ao ync oolefs vth eelsasav,v yain rogers a edxcapapentna sbbivleee
8, 2012 | 0.1021/b achiTevheed 1 f9o8r0 sst usadwie sa r beqooumirin ign rimepmetuitnivoec haenmaliycsails roerse hairgchh saanmdp ldee vloealodpsm. ent
October 6 | doi: 1 wbfoeort rtekeren dvm iwreoitthnhom dsecsni etfanoltr i satimps pmalitcu afnetodiocenhrsae.ml aicRnaedls esaatnaratclehy esrirses.g uaElatnt oturrneypi varegernesnietucyire slsa bbtaoos redadeto vnreeileowsp
32 on 3, 199 businesses on the desirability of immunoassay test kits for site monitoring
6.152.ober 2 nanizdi nhgu tmhea nco esxtp soasvuinreg sa isnse ismsmmeunnto. chMemajoicra lc ahnemalyicsaesl, madandruefsascetdur iesrssu,e rse scuocgh
7.6Oct as the need to quantify cross-reactivity and minimize matrix effects in
oaded by 21ation Date: bdoreudgcaetsrn. ttoIo n lo atwhcceeer p 1tt9h 9ec0 escr,ot asthitn eo fiUm r.emSg.ui sEntrnoacvthiiroeonmn miacnaednl tpraerl-o rPceergdoisutterrceatstii oofnon r Ao tgfh epene csaytni coaidflyfeisc iispa rloloyf
wnlblic several compounds. Back in the laboratories, new and exciting refine
DoPu ments were being discovered. Research and regulatory interest escalated.
The race was on.
In 1992, the U.S. Environmental Protection Agency laboratory in Las
Vegas (at that time the Environmental Monitoring Systems Laboratory,
now the National Exposure Research Laboratory, Characterization
Research Division), seeing the need for intra- and interagency communi
cation, sponsored the first Immunochemistry Summit Meeting. Since
then, the summit has been held annually, growing to an attendance of
about 150 in 1995 at Immunochemistry Summit IV. Summit IV featured
speakers from university research centers, government laboratories,
major chemical companies, and commercial producers of immunochemi
cal reagents, equipment, and immunoassay test kits. Representatives from
local, state, and federal government laboratories participate in the
ix
In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
summits and provide regulatory insight as well as case studies that illus
trate the success or failure of immunochemical methods for a variety of
applications.
This volume is intended to inform novice readers about the strength
and versatility of immunochemical methods for a wide spectrum of analyt
ical environmental applications and to provide experienced researchers
with a selection of readings in various areas of applied environmental
immunochemistry. Authors from regulating agencies present the latest
uses, both accepted and innovative, of immunochemical and related
methods as well as quality assurance guidelines. Authors from universi
ties present updates on breakthrough research. Authors from manufac
turing companies provide insight into recently developed products and
procedures.
org 001 The U.S. Environmental Protection Agency is proud to have initiated
s.pr
ac6. this recognized forum for technical communication in the area of immu
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October 8, 2012 | 6 | doi: 10.1021/b wmftmiioeiaotnlhrydsie , no bf lpe ikr itamehhc eemttl hidtuieo En jnAonoeilvmrnoisgtrel ioyrconia cfmwla intmetrnh aCet dtahhpiltoe rimdooPsnfie.rca soalst Aileo aScnnntoa iacoellil yneettsc iyoct,raAc oilweg nteicitincheh ces yim am'ansimns tedruyNy net aoe itcctniohhotwnenomaia crltai hsdlte Erro yexrce pgfxrooaucrsniiututiiiznrmnaegg
6.152.32 on ober 23, 199 wtRhieetsh es auIrnmcthemr niLte atmb oaerceactietnosgsr.sy tDhooi gmthiteea lpn caeoxgmte m lweuvielnlli cboaeft i oatenvca hwilnaiilbcll aebl r tiinnot get rethcahcetn inoicenat.wl opWrekreisn ohgno nopefe l
7.6Oct that a combined force of publications, conferences, focused work groups,
ded by 21on Date: toahnf ed imdeelmevceutlornoponecrihcse mfaonircduam lu sstee rwcshi lnolo fr lieonsgnuioeltvs a intainv aedn s ace ixecpnoatninftdiince udme daew thcaoordoespn.e ersast ioofn t hbee tpwoewener
oaati
wnlblic
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DP JEANETTE M. VAN EMON
JEFFRE C. JOHNSON
Characterization Research Division
National Exposure Research Laboratory
U.S. Environmental Protection Agency
P.O. Box 93478
Las Vegas, NV 89193-3478
CLARE L. GERLACH
Lockheed Martin Environmental Systems
980 Kelly Johnson Drive
Las Vegas, NV 89119
May 3, 1996
χ
In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
Chapter 1
Environmental Immunochemistry: Responding
to a Spectrum of Analytical Needs
Jeanette M. Van Emon1 and Clare L. Gerlach2
1Characterization Research Division, National Exposure Research
Laboratory, U.S. Environmental Protection Agency, P.O. Box 93478,
Las Vegas, NV 89193-3478
acs.org 6.ch001 2Lockheed Martin EnvirLonams eVnetgaal s,S NysVte m89s1,1 998 0 Kelly Johnson Drive,
October 8, 2012 | http://pubs.6 | doi: 10.1021/bk-1996-064 TboEsiimspfinh oebsmvelceoo iivufrtgaihoeneyl nrotla,aidm aeal bs sneosse,cdn afn ii tyeeeena fnnnlci tvtvtlei iuiifmasrridonotcm innd nskmm kugaiin teel ac lonnosnoctt rwnaaah lllfeiay utemihstsmni icitgocsmhaintoenlru m yetcenoe hxpor cpeclicaneimrhtxngaeeic . msmb attiTeerstiyins h oat,tai rn ssynobe mf i rsb oisiinurncm,i tlnhnretpeioeggdlwmvrseiaa s atittcaosreiidtvsdgprie eylna it,tsgnh eym daecasrihr tosy emnpsl meoneoaclvass outeattugelhrbarryaalyerle.t
32 on 3, 199 pemropcelossy.s Timhem gurnoowcihnegm reigcualla tteocrhyn aiqnude uss ears a ac cceopmtapnocne eonft imofm au mnouclhtiesmteip
7.66.152.October 2 cgarol wmtehth ino dths isfo tre cdhonzoelnosg yof - r eagt uthlaet eladb c boemncpho,u antd tsh ee nhsauzraerds othues wcoansttien usieted,
ed by 21n Date: aaqgnurdain cbtuieflytiucoarnatdilo . nrAu npoopffl fif,co aoatdiso sneassn sdamr epe hnwat rimdoefas chpeuruematidacn,a li napcnuldurdi etiync,og laodnegdtiec rgamrl oieunxnaptdiowosnua troeefr,
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Environmental immunochemistry has grown dramatically in the academic, commer
cial, and regulatory areas over the past fifteen years. Before the 1980s, immunochemi
cal methods were widely used in clinical applications (7) and their success in these
critical studies led environmental scientists to consider immunoassay use for screening
of hazardous compounds in various media. Body fluids are complex media, but new
challenges were presented by soils, sludges, food, and agricultural products. Methods
were developed, test kits were designed and manufactured, and many comparison
studies were initiated to evaluate the performance of these environmental analytical
newcomers.
It is appropriate at this juncture in the development and use of environmental
immunoassays, to review the success of the methods, assess the status of regulatory
0097-6156/96/0646-0002$15.00/0
© 1996 American Chemical Society
In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
1. VAN EMON & GERLACH Responding to a Spectrum of Analytical Needs 3
acceptance, and welcome the next tier of practitioners to the arena of immunochemical
and related technologies. This volume presents in-depth research reports, environmen
tal applications studies, data interpretation subtleties, and commercial success stories.
It is hoped that the scope of this monograph will make it interesting to the experts, give
new applications ideas to researchers, and provide a strong technical basis for novice
users. The multidisciplinary character of immunochemical technology is one of the
strengths of the Immunochemistry Summit Meetings, upon which this volume is
based. The popularity of the Summit Meetings is due to the recognized value of inter
agency and intercorporation exchange of ideas.
All immunoassays are based on the interaction between an antibody and a
target analyte. Antibodies are produced in response to an immunogen by a complex
mechanism (2). Recently, as analytical chemists have become more interested in the
8, 2012 | http://pubs.acs.org 0.1021/bk-1996-0646.ch001 thbbtcRehoaeu.c isyntshEe ot b.nivd nneoB oduntllioo eeuRat gf hgmtteiyehotistnei,e eer pi b da a uasaern tsvcnt(adth5hadet u)i i rtil.fshns a eq e bpClitu lnhiaotaol eeliwn1mptc yta9a himr r9tbneaeo0leiatsfeic.re v mocaaaTefilrt ao h chemltrheh nueme eemv ctra eahosSanlnom onudaudp tmb fes beaitx nhm,echp tteeeauotixd vetrsN t ereMuee arrnanbsteendeti ehoeamiotnunnlifyongsa iidtngl maii eicsmIstmv.namo melsru tl itiounonotfpnugoo eto atleihdscss .esh, f d asaoeeUyenmrs dngctOie ircrtvischoabtecuel e ruk pcdsupisai sttia psenyit sari ht o rbchaaeniovipslsamieep lt ae aicmeSrdrocseaiunh tnfwineteotrite rintiyslny l.
October 6 | doi: 1 taeenxsdctr Hektiietosan lft ohor f( NmtoIeOxaisScuH rci)on mpgr peaosleauncnhtd lwso roi srm kre etthlaiabatno dtl ietuempso oinnn s sutreravitneeersa. tlh Afea bcestfoofirrcpsi teianoncndy, ptohafir sitm imtimounultininvogaa,rs aisanatdye
32 on 3, 199 character presents a challenge in data interpretation as well as in analytical procedures.
6.152.ober 2 Eanxtaibmopdlieess oafn tdh ea nretisbeoadrcyh teact hNnIiqOuSeHs atore mproensietnotre de,x pinocsulurde ining tthhee uusrein oe f ocfi recxuplaotsinedg
7.6Oct workers. Research at NIOSH demonstrated that circulating antibodies to morphine can
ded by 21on Date: bbsaeey p4 tr-ee5ss tet inkmitt ,ei nso trmhigeo iraneba slsleyenn cdseeit voivef eluo ripnien ddar efytoe arc ntaianllagyct hethlsoe. r Ipanrn iaamnlyaosrtheyse h ri unNm gIaOronSu Hmnd eswttauabdtoeylr,i, t aewn oaimfs afmolauucnnhodloa tsro,
ownloaublicati aimlamchulnoor amssearyc atpetcuhrnaitqeu, eths aanr ei no rddeetersc toinf gm tahgen pitaurdeen tm moorele sceunlesi. tiFveo rt hsaonm ter acdoimtiopnoauln gdass,
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chromatographic/mass spectrometric techniques. Benefits such as increased sample
throughput, reduced cost, simpler sample preparation and no derivitization steps make
this type of analysis very attractive.
V. Lopez-Avila and J. M. Van Emon discuss their work in the coupling of
immunoaffinity chromatography (IAC) extraction with on-line liquid chromatography
and mass spectrometry (MS). The IAC technique is based on the ability of antibodies
to separate a target analyte from the complex matrices that often challenge environ
mental analytical chemists. On-line analysis is done by high-performance liquid
chromatography (HPLC) and electrospray MS. The system is particularly useful for
the analysis of compounds that are water-soluble, nonvolatile, thermally labile, or
highly polar. Sample throughput of the MS is increased by integrating IAC with the
analytical instrumentation, providing automated, streamlined sample preparation.
In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
4 ENVIRONMENTAL IMMUNOCHEMICAL METHODS
I. Wengatz and co-workers at the University of California-Davis are developing
immunoassays to assess human exposure to xenobiotics, such as pesticides (4).
Metabolites of certain xenobiotics may serve as biomarkers of exposure in toxicity
studies. The UC-Davis group is developing immunoassays for trace levels of triazine
herbicides and their metabolites, and is using cross-reactivity information to enable
antibody use for screening classes of analytes. Human exposure research is increasing
ly important as regulatory agencies move from prescriptive methods based on absolute
contaminant concentration to risk-driven guidelines based on the bioavailability of the
contaminant and its threat to human and ecological health.
Other immunochemical-based technologies, such as biosensors, are being de
veloped at the Naval Research Laboratory. L. Shriver-Lake and fellow researchers
describe a continuous flow immunosensor that can be used to measure small molecules
8, 2012 | http://pubs.acs.org 0.1021/bk-1996-0646.ch001 fbioiaasnmiimdanb gsvd menpeaairaesd u,ncc lr nt.oiroo asoemmgn taaee Eespcl stssolesr aoeoaintycci fmdbt asticure lvpolociwlyenncet .ivigshi ntm eO ehgosnm mt. rt eut Sihiiulodcnaeenniadc emoaltied rklaon io.s im pznmasrRyacimomtymteoviu vsurdebnpiene neoo osapgian ancoa sspstgs leissrya va otpyamiiycesttsée u re mmsfssro sas eeb rrr.taemse ehtsp Iirouemnoebddrargaesmt m s ,doo deu sanndenb.s t oty eehwAsol reetemneh c nfl eftilsirin m boboaceerescohdsr rhad t ohenpcipmfpegoroi toecritioec vaecp iatn inobidtltoi fiietoiono hm antssmeh e ml oenoeoff utsnlaf ruoon ngiobcroe a,rtaons laeelybzpislnatynytcisismi eccoenonaardegrltsl
October 6 | doi: 1 rdeeaslc-rtiibmeTedh femo pro rntohimteo iarsinen agolf y msthiesea sosefu mpreoemltyheconhdtslso. riisn Aiantne t dhe ebl eiepcvthereonnctuyhaleslm . diecvael loimpmmeunnto osef nas soern sios r atlhsaot
32 on 3, 199 can be used remotely, gathering information without an operator and sparing personnel
6.152.ober 2 the possSib. lCe oexupltoesru raen da sasosscoiactieadte sw idthis csoumsse ae nsvoilriodn mstaetnet asly wstoermk. that combines the
7.6Oct advantages of optical sensing and competitive immunoassays. This sensing package
ed by 21n Date: cchoemmpirsistreys, aa nlidg htht es oauprpcreo pwrhiaitceh dpertoevcitdore.s tThhei so ucthpiupt- btharsoeudg she nthsoer w isa veeagsiulyid em, athneu fsaecntusirnedg
do and has a sensing arm and a reference arm. By combining fiber optic chemical sensor
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wnlblic technology with immunoassay, these systems enlarge the panorama of analytical tools
ou available to environmental scientists. Research that reduces the number of steps in an
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analytical procedure will be appreciated as the environmental analysis emphasis moves
from the laboratory to the hazardous waste site.
Immunoassay test kits and other immunochemical procedures are now used
almost routinely to monitor the purity of food and drugs, and drinking water.
Immunoassay test kits are increasingly used at hazardous waste sites regulated under
Comprehensive Environmental Response, Compensation and Liability Act (CERCLA)
and Resource Conservation and Recovery Act (RCRA). An important step was taken
in 1993 when the EPA's Office of Solid Waste and Emergency Response (OSWER)
accepted nine immunoassay methods for its compendium, SW-846 (5). This
regulatory acceptance means that these methods can be used for certain RCRA
applications. The EPA's Office of Water and Office of Drinking Water are utilizing
field methods such as immunoassays to determine the safety of the nation's water
supplies for drinking, agriculture, and recreational use (6). The U.S. Geological
In Environmental Immunochemical Methods; Van Emon, J., et al.;
ACS Symposium Series; American Chemical Society: Washington, DC, 1996.
