Trace EnvironmentalQuantitative AnalysisPrinciples, Techniques,and ApplicationsSECONDEDI TI ON 2006 by Taylor & Francis Group, LLCA CRC title, part of the Taylor & Francis imprint, a member of theTaylor & Francis Group, the academic division of T&F Informa plc.Trace EnvironmentalQuantitative AnalysisPrinciples, Techniques,and ApplicationsSECONDEDI TI ONPaul R. LocontoBoca Raton London New York 2006 by Taylor & Francis Group, LLCPublished in 2006 byCRC PressTaylor & Francis Group 6000 Broken Sound Parkway NW, Suite 300Boca Raton, FL 33487-2742 2006 by Taylor & Francis Group, LLCCRC Press is an imprint of Taylor & Francis GroupNo claim to original U.S. Government worksPrinted in the United States of America on acid-free paper10 9 8 7 6 5 4 3 2 1International Standard Book Number-10: 0-8247-5853-6 (Hardcover) International Standard Book Number-13: 978-0-8247-5853-0 (Hardcover) Library of Congress Card Number 2005048512Thisbookcontainsinformationobtainedfromauthenticandhighlyregardedsources.Reprintedmaterialisquotedwithpermission,andsourcesareindicated. Awidevarietyofreferencesarelisted.Reasonableeffortshavebeenmadetopublishreliabledataandinformation,buttheauthorandthepublishercannotassumeresponsibility for the validity of all materials or for the consequences of their use.Nopartofthisbookmaybereprinted,reproduced,transmitted,orutilizedinanyformbyanyelectronic,mechanical,orothermeans,nowknownorhereafterinvented,includingphotocopying,microlming,andrecording, or in any information storage or retrieval system, without written permission from the publishers. Danvers, MA 01923, 978-750-8400. CCC is a not-for-prot organization that provides licenses and registrationforavarietyofusers.FororganizationsthathavebeengrantedaphotocopylicensebytheCCC,aseparatesystem of payment has been arranged. Trademark Notice: Product or corporate names may be trademarks or registered trademarks, and are used onlyfor identication and explanation without intent to infringe.Library of Congress Cataloging-in-Publication DataLoconto, Paul R., 1947-Trace environmental quantitative analysis : principles, techniques, and applications / PaulR. Loconto.--2nd ed.p. cm. Includes bibliographical references and index.ISBN 0-8247-5853-6 (alk. paper)1. Environmental chemistry. 2. Trace analysis. 3. Chemistry, Analytic--Quantiative I.Title.TD193.L63 2005628.50287--dc22 2005048512Visit the Taylor & Francis Web site at and the CRC Press Web site at Taylor & Francis Group is the Academic Division of T&F Informa plc. 2006 by Taylor & Francis Group, LLCForpermissiontophotocopyorusematerialelectronicallyfromthiswork,pleaseaccesswww.copyright.com(http://www.copyright.com/)orcontacttheCopyrightClearanceCenter,Inc.(CCC)222RosewoodDrive,http://www.taylorandfrancis.comhttp://www.crcpress.comDedicationTo my ve points of light. Each added a new dimension to my life.Jennifer AnnMichelle AnnAllison MarieJulia MarieElizabeth MarieIn memory of Taylor Renee Hamel(19952005)whose light was extinguishedmuch too early. 2006 by Taylor & Francis Group, LLCPreface to the Second EditionTherapidpaceinwhichtraceanalysisischanginghaswarrantedthewritingofasecond edition in a relatively short period. What is new? The second edition attemptstomovethereaderfromthemostelementaryofprinciplesoftraceenvironmentalquantitativeanalysis(TEQA)tothosetechniquesandapplicationscurrentlybeingpracticedinanalyticallaboratoriesdedicatedtotraceenvironmentalchemicalandtrace environmental health quantitative analysis while adding new signicant topics.Theincreasingimportanceofmassspectrometrywillbecomeapparenttothereader primarily as a low-resolution hyphenated technique. The principles that under-determinative techniques.alternativestoliquidliquidextractionareintroduced.Columnchromatographiccleanup,virtuallyignoredearlier,andgelpermeationchromatographyhavebeenintroduced along with additional applications to biological sample matrices of envi-ronmental health and toxicological interest. Matrix solid-phase dispersion as appliedtotheisolationandrecoveryofpersistentorganicpollutantsfromshtissuehasbeen added. The prolic growth of SPME as evident in the analytical literature overthe past 5 years has warranted an enlarged section on this technique.More than two dozen new topics not previously discussed in the original bookhave been added to the second edition.Anyauthor,uponreviewingthenishedproductofarstbook,hasamostimmediate desire to rewrite all of it. I have resisted this temptation and have modiedonly those sections of the original book that I felt enlarge and enhance the environ-mental analytical chemists understanding of TEQA.Who should read the second edition? Scientists, in addition to analytical chem-ists,suchasorganicchemists,biochemists,molecularbiologists,geologists,toxi-cologists,epidemiologists,foodscientists,andchemicalandenvironmentalengi-neers will nd that the second edition might enhance their understanding of TEQA.Laboratorytechniciansofvariousskillandknowledgelevelsshouldalsondthecontent of this edition benecial.Thestyleforthesecondeditionhasremainedthesame.Sectionheadingscontinue to be cast in the form of a question. New terms have been italicized whentheyappearforthersttime.BeneatheachchaptertitleisabriefChapterataGlance so that the reader can more quickly nd topics of immediate interest. Figuresandtablesarebothseparatedandnumberedinsequenceandintegratedinthetextwithout numbering, as before. Digressions from the main topics have also occurredinamannersimilartothatintheoriginalbook.GraphsareeithersketchesthatIdrew to illustrate a principle or carefully drawn from experimental data (Im a prettygood chemist; an artist, I am not). To reiterate from the preface to the original book, 2006 by Taylor & Francis Group, LLClie GC-MS, GC-MS-MS, LC-MS, and ICP-MS can now be found in Chapter 4 onChapter 3 on sample preparation techniques has been enlarged so that even moreIhavetriedveryhardtomakethistextreadable,interesting,andrelevant,andatthe same time, introduce sound principles and practices of TEQA.I express my gratitude to the Division of Chemistry and Toxicology, Bureau ofLaboratories,MichiganDepartmentofCommunityHealth;theMichiganPublicHealthInstitute;andtheBiomonitoringPlanningGrant,NationalCenterforEnvi-ronmentalHealth,CentersforDiseaseControlandPrevention.Theseinstitutionsand grants enabled me to nd the time to write, edit, and rewrite. Barbara MathieuandcolleaguesatTaylor&Francishavepainstakingly,forasecondtime,turnedthis authors rough draft into a book. My wife, Priscilla, has graciously endured herhusbands passion for writing. And my motivation to write is rooted in and summa-rized by the ancient Chinese Proverb:I hear and forget; I see and I remember; I write and I understand. 2006 by Taylor & Francis Group, LLCAbout the AuthorPaulR.LocontoiscurrentlyalaboratoryscientistspecialistwiththeMichiganDepartmentofCommunityHealth,BureauofLaboratories,Lansing.Dr.Locontois the author of 24 peer-reviewed publications and 33 oral and poster presentationsintraceanalysisandchemicaleducation.Hecombinesvariousworkexperiencesthat include teaching at a community college, managing an environmental engineer-ing research laboratory within a large university, and conducting analytical methoddevelopmentforanindependentenvironmentaltestinglaboratory.Allhavegiventhe author many unique insights over the years into the principles, techniques, andapplications of TEQA. 2006 by Taylor & Francis Group, LLCTable of Contents1. Introduction to Trace Environmental Quantitative Analysis (TEQA) 12. Calibration, Verication, Statistical Treatment of Analytical Data, Detection Limits, and Quality Assurance/Quality Control 373. Sample Preparation Techniques to Isolate and Recover Organics and Inorganics 1194. Determinative Techniques to Measure Organics and Inorganics 3235. Specic Laboratory Experiments 547Appendix A: Glossary 651Appendix B: QA/QC Illustrated 679Appendix C: TEQA Applied to Drinking Water/Computer Programs for TEQA 689Appendix D: Instrument Designs 705Appendix E: Useful Internet Links for Environmental Analytical Chemists 711Appendix F: Useful Potpourri for Environmental Analytical Chemists 717 2006 by Taylor & Francis Group, LLC11Introduction to Trace Environmental Quantitative Analysis (TEQA)Ifyouteachapersonwhattolearn,youarepreparinghimforthepast.Ifyouteachhim how to learn, you are preparing him for the future.AnonymousCHAPTER AT A GLANCECase study from trace enviro-health quantitative analysis ....................................... 2Case study from trace enviro-chemical quantitative analysis................................... 4Extent of chemical contaminants in humans ............................................................ 7Analytical chemistry approaches to biomonitoring................................................ 11Environmental chemistry......................................................................................... 11EPA regulations ....................................................................................................... 15Analytical methods that satisfy EPA regulations.................................................... 20Physical/chemical basis for EPAs methods protocols............................................ 32References................................................................................................................ 35Asweapproachedthenewmillennium,thenewsmediaandrelatedmassmediaspeculated on what would be different. The 20th century was gone. The 21st centurywas upon us. The tragic events of 9/11 in the U.S. provided one such answer. Since9/11, questions such as If we have a terrorist event, can we measure trace concen-tration levels of terrorist-related chemical substances and attempt to evaluate expo-sure over relatively large numbers in the population? have shifted the dialogue.Public health laboratories are beginning to respond to this terrorist-related threat.These laboratories are moving toward having a capability in trace environmental healthquantitativeanalysis(alsoabbreviatedTEQA).Atthesametime,biomonitoring-relatedinitiativesareexpanding.FederallaboratoriessuchastheNationalCenterforEnvironmentalHealthandtheCentersforDiseaseControlandPrevention(NCEH/CDC) are assisting state labs in the transfer of both bioterroism- and biomon-itoring-relatedanalyticalmethods.Thesemethodsaredesignedtomeasuretrace 2006 by Taylor & Francis Group, LLC2 Trace Environmental Quantitative Analysis, Second Editionconcentrationlevelsofchemicalsubstancesthateitherpersist(persistentorganicpollutants(POPs))orareeliminatedratherquicklybythebody,i.e.,nonpersistentorganic pollutants (NPOPs).Bioterrorism and biomonitoring are key initiatives that are currently driving thechanging nature of trace quantitative organics and inorganics analysis. The secondeditionofthisbookattemptstoreectthesechanges.Thisnewemphasis,whenanalysis, has led this author to adopt a new term: trace enviro-chemical/enviro-healthquantitativeanalysis,whoseacronymisalsoTEQA.Ihavetriedtoaddthoseanalytical concepts that are most relevant to conducting trace enviro-health quanti-tativeanalysis.Sampling,samplepreparation,determinativetechnique,anddatareduction/interpretationareverysimilartobothtraceenviro-chemicalandtracethe enviro-health aspects of trace quantitative organics and inorganics analysis whilediscerning the similarities and differences in both. One starts with an understandingof the chemical nature of the sample or human or animal specimen received. A clientneedstounderstandjustwhatanalytesaretobemeasuredandhowthesetwopathways lead to four steps in the process shown in Scheme 1.1. There is no substitutefor effective communication between the client and the analytical laboratory. Sam-determinativetechniques,oftenreferredtoasinstrumentalanalysis(introducedindata (introduced in Chapter 2) comprise the important aspects of successfully imple-menting TEQA.Thissecondeditionintroducesprinciplesandpracticesoftraceenviro-healthquantitativeanalysiswhileexpandingontheprevioustreatmentoftraceenviro-chemical quantitative analysis where the emphasis was placed only on environmentalsamples.1TwocasestudiesdrawnfromtherecentliteratureintroducethepracticeofcontemporaryTEQA.Therstcasestudydemonstratesthatapossibleendocrinedisrupter can be isolated and recovered from human urine.1. CAN AN EXAMPLE PROVIDE INSIGHT INTO TRACE ENVIRO-HEALTH QA?Yes,westartbybrieyintroducingresultsfromapublishedstudy.BisphenolA(BPA), 2,2-bis(4-hydroxyphenyl)propane, is an industrial chemical used in a varietyof plastic materials, some of which are used in packaging, and hence is believed toleach out into consumable items such as food and dental llings. One way to betterassess human exposure to BPA despite the source of exposure being largely unknownis to biomonitor, i.e., to measure, trace BPA in human urine. Brock and coworkers2attheNCEH/CDChavedevelopedaquantitativeanalyticalmethodtodeterminejusthowmuchBPAmightbepresentinaurinesampleobtainedfromapersonbelievedtobeexposedtoBPA.2BPAisapparentlyexcretedeitherunmetabolized 2006 by Taylor & Francis Group, LLCcombinedwiththemoreestablishedmethodsoftraceenvironmentalquantitativeenviro-health quantitative analysis. Scheme 1.1 depicts both the enviro-chemical andpling(introducedinChapter2),samplepreparation(introducedinChapter3),Chapter 4), and data reduction, statistical treatment, and interpretation of analyticalIntroduction to Trace Environmental Quantitative Analysis (TEQA) 3or glucuronidated. Urinary BPA glucuronide seems to be a longer-lived biomarker(12 to 48 h). After deglucuronidation using -glucuronidase, BPA was isolated andrecovered by reversed-phase solid-phase extraction. The isolate was converted to itspentauorobenzyl ether. The pentauororbenzyl ether of BPA was quantitated usingisotope dilution gas chromatography-mass spectrometry (GC-MS). A method detec-two different mass spectra for the pentauorobenzyl ether of BPA that eluted fromthe gas chromatographic column at 26.4 min. The top mass spectrum in Figure 1.1was obtained via electron-impact mass spectrometry and reects positive fragmentions, while the bottom mass spectrum was obtained via negative chemical ionizationmass spectrometry. Pooled human urine samples showed no detectable BPA beforethe urine was treated, while BPA concentration levels varied from 0.11 to 0.51 partsSCHEME 1.1What is thechemical natureof the sample orhuman specimen?Blood, plasma,serum, urine,breast milk,adipose tissue,saliva, otherbody uids Air, groundwater, surfacewater,wastewater,plant euent,leachate,soil, sediment,y ash,biomass,otherenvironment-al matricesTrace Enviro-chemical quantitative analysisPriority pollutants asanalyte(s) of interestPersistent or non-persistent organicpollutants as analyte(s)of interestApplication of appropriatedeterminative techniques(instrumental analysis)Data reduction, statisticaltreatment of analytical data andinterpretation; detection limitcalculations; QA/QCReporting ofanalytical resultsto clientSampling, samplepreparationTrace Enviro-healthquantitative analysis 2006 by Taylor & Francis Group, LLCtionlimit(MDL)wasreportedtobe120partspertrillion(ppt).Figure1.1shows4 Trace Environmental Quantitative Analysis, Second Editionper billion (ppb) for the treated urine. Molecular structures for Bisphenol A and forpentauorobenzyl bromide (-bromo-2,3,4,5,6-pentauorotoluene) are shown below:Thesecondcasestudydemonstratesthatanemergingpharmaceuticalcanbeisolated and recovered from wastewater.2. CAN AN EXAMPLE PROVIDE INSIGHT TO TRACE ENVIRO-CHEMICAL QA?Yes indeed, and we start with a published report on the isolation and recovery of clobricacidfromwastewater.3Clobricacid[2-(4-chlorophenoxy)-2-methyl-propanoic]FIGURE 1.1 Electron impact (top) and negative chemical ionization (bottom) mass spectraof the pentauorobenzyl ether of Bisphenol A.573 (M-CH3)+10080604020Relative abundance700 600 500 400 300 200299588 (M)+EI+211FFFF FOF FFF FO10080604020 Relative abundance650 400408407350 500m/z450 600 550NCIF FFF FOOOO HH BisphenolA BrF F F F F Pentauorobenzyl bromide 2006 by Taylor & Francis Group, LLCIntroduction to Trace Environmental Quantitative Analysis (TEQA) 5acid is the bioactive metabolite of various lipid-regulating prodrugs. Acidic metab-olites of pharmaceuticals present one type of analyte that appears in the efuent ofmany municipal treatment facilities. The isolation and recovery of clobric acid isconsistent with the Environmental Protection Agencys (EPA) Division of Environ-mentalSciences,EnvironmentalChemistryBranchsmissiontostudythefateandtransportofchemicalcompoundsderivedfrompharmaceuticals,theirmetabolites,and personal care products. Patterson and Brumley approached the need to quantitateclobricacidbycomparingtwomajortypesofsamplepreparation,liquidliquidextraction (LLE) and reversed-phase solid-phase extraction (RP-SPE), using a sty-rene/divinyl benzene adsorbent. The determinative technique used was electron-impactgaschromatography-massspectrometry(EI-GC-MS)afterconversionofclobricacid to its methyl ester by derivatizing with trimethyl silyl diazomethane. An internala trace quantitative analysis of samples of sewage efuent to determine how muchclobric acid is present. Shown below are the molecular structures for clobric acidand two organic compounds, 3,4-D and PCB 104 (2,2,4,6,6-pentachlorobiphenyl),used by the authors to calibrate the instrument based on the internal standard mode:Since EI-GC-MS was the only instrumental determinative technique (determina-subtractedstandardorclobricacidmethylester,whilethemassspectrumshownbelowisforabackground-subtractedmassspectrumobtainedfromtheefuentClobric acid 3, 4-DClClOOOHClOOOHClClClClCl2, 2', 4, 6, 6'-Pentachlorobiphenyl 2006 by Taylor & Francis Group, LLCin Figure 1.2. The mass spectrum shown on top in Figure 1.2 is for a background-standard mode of instrument calibration (introduced in Chapter 2) was used to providetive techniques are introduced in Chapter 4) used, two EI mass spectra are compared6 Trace Environmental Quantitative Analysis, Second Editionsampleextractattheretentiontimeofclobricacidmethylester.Thedisputablefactthatbothmassspectraareidenticaldemonstratestheunequivocalnatureofidentication,sometimesreferredtoinEPAmethodsasconrmation.Figure1.2illustrates trace environmental qualitative analysis. Using all abundant fragment ionsor even one or more selected fragment ions with which to build a calibration curve,and from this curve to interpolate and thus to nd how much clobric acid is presentin the unknown extract from the waste efuent, nicely illustrates the science of traceenvironmental quantitative analysis.Let us summarize some regulatory issues, rst from this emerging enviro-healtharena.Wethencompletethisintroductorychapterwithanemphasisonthewell-establishedenviro-chemicalarena,largelyreviewingthesignicantenvironmentalregulations. We then show just how the EPA methods t in. A signicant questionis before us with respect to enviro-health.FIGURE 1.2 EI mass spectra for clobric acid methyl ester.10080604020Rel abundance127.128130169154.m/z181.10080604020Rel abundance127.128130169154.m/z181. 2006 by Taylor & Francis Group, LLCIntroduction to Trace Environmental Quantitative Analysis (TEQA) 73. TO WHAT EXTENT DO ENVIRONMENTAL CONTAMINANTS ENTER HUMANS?therstNationalReportonHumanExposuretoEnvironmentalChemicals,con-ducted by the CDC. This report provides exposure information about people partic-ipating in an ongoing national survey of the general U.S. population the NationalHealthandNutritionExaminationSurvey(NHANES).ThesurveywasconductedbytheNationalCenterforHealthStatisticsoftheCDC.ThisrstreportpresentsdataforthegeneralU.S.populationfromthe1999NHANES.Accordingtothereport,thissurveywasconductedinonly12locationsacrossthecountry.Mostanalyseswereconductedinsubsamplesforthepopulation.Moredatawouldbeneededtoconrmthesendingsandtoallowmoredetailedanalysistodescribeexposure levels in population subgroups.4All the metals determined are listed in Table 1.1, while just those organics thatreveal a level above the limit of detection are shown in Table 1.2. The report makesTABLE 1.1Geometric Mean of Blood and Urine Levels of Environmental MetalsMetalHuman SpecimenNo. of People Sampled UnitsGeometric Mean (95% Condence Interval)Cadmium Blood 3189 g/L aLead Blood 3189 g/dL 1.6 (1.41.8)Mercury in children 15 years Blood 248 g/L 0.3 (0.20.4)Mercury in females 1649 years Blood 679 g/L 1.2 (0.91.6)Antimony Urine 912 g/L 0.1 (0.090.12)Barium Urine 779 g/L 1.6 (1.51.7)Beryllium Urine 1007 g/L a Cadmium Urine 1007 g/L 0.32 (0.300.33)Cesium Urine 1006 g/L 4.7 (4.25.2)Cobalt Urine 1007 g/L 0.36 (0.360.40)Lead Urine 1007 g/dL 0.80 (0.680.91)Molybdenum Urine 904 g/L 48.4 (43.653.2)Platinum Urine 1007 g/L a Thallium Urine 974 g/L 0.19 (0.170.20)Tungsten Urine 892 g/L 0.10 (0.090.12)Uranium Urine 1006 g/L 0.008 (0.0060.001)aNot calculated; the proportion of results below the limit of detection was too high to provide a validresult.Source: Adapted from the National Health and Nutrition Examination Survey (NHANES), U.S., 1999.CDC,NationalReportonHumanExposuretoEnvironmentalChemicals,CDC, Atlanta,GA,March2001. 2006 by Taylor & Francis Group, LLCTable 1.1 (metals) and Table 1.2 (organics) highlight selected analytical results from8 Trace Environmental Quantitative Analysis, Second Editionitveryclearthatthepresenceofdetectableconcentrationlevelsofchemicalsub-stancesdoesnotindicatethatthechemicalcausesdisease.Since1976,CDChasthat the geometric mean blood Pb levels for children aged 1 to 5 have decreased to2.0 from 2.70 g/dL, the geometric mean for the period 19911994. These decreasesin blood Pb levels indicate a success in public health efforts to decrease the exposureof children to Pb.Not shown in either Table 1.1 or Table 1.2 are the results for reduced exposureof the U.S. population to environmental tobacco smoke (ETS). Cotinine is a metab-olite of nicotine that tracks exposure to ETS. Molecular structures for both cotinineand its precursor, nicotine, are shown below:Adecreaseinserumcotinineconcentrationlevelsfrom0.20ng/mLobtainedduring the period 19881991 to 0.050 ng/mL (obtained in this study) among peopleaged 3 years and older (a 75% decrease) indicates a dramatic reduction in exposureof the general population to ETS over the past decade.Table 1.2 reveals some surprising results. CDC scientists measured metabolitesof seven major phthalates. Di-2-ethylhexyl phthalate and di-iso-nonyl phthalate aretwophthalatesproducedingreatestquantity;however,metabolitesofdiethylandTABLE 1.2Geometric Mean of Blood and Urine Levels of Environmental OrganicsOrganic Pesticide or MetaboliteHuman SpecimenNo. of People Sampled UnitsGeometric Mean (95% Condence Interval)Dimethyl phosphate Blood 703 g/L 1.84 (1.102.59)Dimethyl thiophosphate Blood 703 g/L 2.61 (1.773.45)Diethyl phosphate Blood 703 g/L 2.55 (1.333.78)Diethyl thiophosphate Blood 703 g/L 0.81 (0.690.94)Dimethyl dithiophosphate Blood 703 g/L 0.51 (0.390.62)Diethyl dithiophosphate Blood 703 g/L 0.19 (0.140.23)Monobenzyl phthalate Blood 1029 g/L 17.4 (14.120.7)Monobutyl phthalate Blood 1029 g/L 26.7 (23.929.4)Monoethyl phthalate Blood 1029 g/L 176.0 (132220)Mono-2-ethyl hexyl phthalate Blood 1029 g/L 3.5 (3.04.0)Source: AdaptedfromtheNationalHealthandNutritionExaminationSurvey(NHANES),U.S.,1999. CDC, National Report on Human Exposure to Environmental Chemicals, CDC, Atlanta, GA,March 2001.NNNicotineNNOCotinine 2006 by Taylor & Francis Group, LLCmeasured blood Pb levels as part of NHANES. Results presented in Table 1.1 showIntroduction to Trace Environmental Quantitative Analysis (TEQA) 9dibutyl phthalate were much higher in the population than levels of metabolites ofthe most ubiquitous phthalates found in the environment.Traceenviro-healthquantitativeanalysis,alsoabbreviatedTEQA,is,inthisauthors opinion, an evolving subdiscipline of trace environmental quantitative anal-ysis.TheClinicalLaboratoryImprovementActof1988(CLIA88)regulatesthechemicallaboratoryandaddressesthoseaspectsoftraditionalclinicalchemistry,such as determining the concentration of creatinine in blood. Toxicological chemistryalsoincludesbloodalcohol,digoxin,lithium,primidone,andtheophyllineassays.The concentrations in the blood and urine of these analytes are signicantly higherthanthosethatwouldbeconsideredatatracelevel.Ourfocusinthisbookistodiscusshowenvironmentalpollutantscanbequantitativelydeterminedinhumanspecimens.However,environmentalprioritypollutantsfoundinhumanspecimensmay have entered the human domain via the various routes of exposure. Figure 1.3FIGURE 1.3 Routes of human exposure to environmental contaminants. Types of body uidsas human specimens for biomonitoring.RespiratorytractInhalationAbsorptionExhalationExfoliationBloodSalivaKidneyIngestionGI tractSkinSweatFecesDevelopingorgansLiverHairUrine 2006 by Taylor & Francis Group, LLC10 Trace Environmental Quantitative Analysis, Second Editiondepicts routes of exposure to environmental priority pollutants along with the pos-sible kinds of body uids, shown as ovals, that could be dened as suitable humanspecimensforbiomonitoring.5Threeroutesofexposureincludeinhalationtotherespiratorytract,ingestiontothegastrointestinaltract,andabsorptionthroughtheskin, often termed dermal exposure. The development of so-called biological markers(biomarkers)representsaveryactiveresearchareainvolvingtoxicologistsandepidemiologists. TEQA has a vital role to play in this research today. A biomarkercanbeeithercellular,biochemical,ormolecularinnatureandcanbemeasuredanalytically in biological media such as tissues, cells, or uids. A suitable biomarkercould be an exogenous substance or its metabolite. It could also be a product of aninteractionbetweenthexenobioticagentandsometargetmolecule.Exposureanddose are two terms that are further elaborated below: Exposure is contact of a biological, chemical, or physical agent with thesurface of the human body. Dose is the time integral of the concentration of the toxicologically activeform of the agent at the biological target tissue. Dose links exposure to risk of disease. Exposure dose effect.The relationship between exposure, dose, and potential health effects is summa-rized below:bloodsupply.Thismodeldoesnotincludeadiposeorotherhumantissue.Bloodandurineareemergingasthemostconvenienthumanspecimenstocollectandconduct biomonitoring.SourcePesticide useAir pollutionWater pollutionConcentration: :Indoor airOutdoor air SurfacesSoilFoodDrinking waterPotentialadverse healtheectsExposurePersonal airDietDermal rinseDoseBiomarkers (urine,blood, hair) 2006 by Taylor & Francis Group, LLCFigure 1.3 depicts a biomonitoring scenario centered with respect to a personsIntroduction to Trace Environmental Quantitative Analysis (TEQA) 11Let us return to the concept of a biomarker as a key ingredient in biomonitoring.Biomarkers provide evidence of both exposure and uptake. The concentration levelofagivenbiomarkerisdirectlyrelatedtotissuedose.BiomarkersaccountforallFactors that limit the usefulness of biomarkers include:5 The fact that many biomarkers are still being developed The need for standardized protocols in both collection and analysis Variability in relationship with exposure Timing each biomarker has a characteristic half-life Expense Difculty in interpreting4. WHAT MIGHT AN ANALYTICAL CHEMISTRY APPROACH TO BIOMONITORING LOOK LIKE?human specimen to analytical result is listed in terms of ve essential and sequentialsteps,eachlinkedbyachain-of-custodyprotocol.Thearrowsshowthattherela-tionship between steps must include a chain-of-custody protocol. This protocol mighttaketheformofawrittendocument.If,however,aLaboratoryInformationMan-agement System (LIMS) is in place, the protocol takes the form of an entry into acomputerthatutilizesaLIMS.ReferringtoFigure1.4,thesamplepreplabmaygive to the analyst a complete sample extract along with a signed chain-of-custodyformtoprovideevidenceastowheretheextractisheadednext.Thisve-stepapproachtobiomonitoringisalsoapplicabletotraceenviro-chemicalquantitativeanalysis.Weleaveforthemomenttraceenviro-healthquantitativeanalysisandpickuptraceenviro-chemicalquantitativeanalysis.Letusrstdenewhatwemeanbyenvironmental chemistry.5. WHAT KIND OF CHEMISTRY IS THIS?The academic discipline of environmental chemistry is a relatively recent develop-ment.Environmentalchemistrycanbedenedasasystematicstudyofthenatureofmatterthatexistsintheair,water,soil,andbiomass.Thisdenitioncouldbeextendedtotheplantandanimaldomainswherechemicalsfromtheenvironmentare likely to be found. This discipline, which developed in the late 1960s, requiresthe knowledge of the traditional branches of organic, inorganic, physical, and ana-lytical chemistry. Environmental chemistry is linked to biotechnology as well as tochemical, environmental, and agricultural engineering practices.Environmental analytical chemistry can be further dened as a systematic studythat seeks to answer two fundamental questions: What and how much matter existsintheair,water,soil,andbiomass?Thisdenitioncouldalsobeextendedtotheplantandanimaldomainsjustdiscussed.Thisdiscipline,whichdevelopedinthe 2006 by Taylor & Francis Group, LLCpossible routes, as shown in Figure 1.3. Biomarkers account for differences in people.OnesuchanswertothisquestioncanbefoundinFigure1.4.Thescenariofrom12 Trace Environmental Quantitative Analysis, Second Edition1970s, spearheaded by the rst Earth Day in 1970 and the establishment of the U.S.EPA, requires a knowledge of traditional quantitative analysis, contemporary instru-mental analysis, and selected topics, such as statistics, electronics, computer software,and experimental skill. Environmental analytical chemistry represents the fundamen-talmeasurementsciencetobiotechnologyandtochemical,environmental,andagriculturalengineeringpractices.Thatportionofenvironmentalanalyticalchem-istry devoted to rigorously quantifying the extent to which chemical substances havecontaminated the air, water, soil, and biomass is the subject of this book.Initsbroadestsense,environmentalchemistrymightbeconsideredtoincludethechemistryofeverythingoutsideofthesyntheticchemistsask.Themomentthatachemicalsubstanceisreleasedtotheenvironment,itsphysico-chemicalFIGURE 1.4 From human specimen to analytical result; the analytical approach to biomon-itoring. Sampling or human specimen collection Sample/specimen preservation and storageRefrigerationAddition of preservativesHolding time considerationsArchive unused specimens Sample/specimen preparation which includes 1. Addition to sample, prior to extraction, of surrogates, labeled isotopes, and internalstandards2. Extraction of analyte(s) of interest frommatrix3. Cleanup of matrix interferences4. Concentration of sample extract 5. Addition of internal standard prior to injection of the sample extract Optimization of determinative techniques and application of quantitative instrumental analysis and includes:6. Calibration and least squares regression using an isotope dilution or internal standardmode of instrument calibration 7. Instrument calibration verication8. Interpolation of the calibration applied tosample extracts for all sample/specimensand QC samples Data reduction and interpretation of analytical data; evaluation of percent recoveries, determining the instrument and method decision and detection limits; statistical treatment of replicate data Implementation of a QA/QC protocol; writing of a QA document that addresses CLIA88 guidelines Implementation of reporting protocols Preparation of summaries, spreadsheets, data bases Archival protocols 2006 by Taylor & Francis Group, LLCIntroduction to Trace Environmental Quantitative Analysis (TEQA) 13properties may have an enormous impact on ecological systems, including humans.Researchers have identied 51 synthetic chemicals that disrupt the endocrine system.Hormone disrupters include some of the 209 polychlorinated biphenyls (PCBs) andsomeofthe75dioxinsand135furansthathaveamyriadofdocumentedeffects(p. 81).6 The latter half of the 20th century has witnessed more synthetic chemicalproductionthananyotherperiodinworldhistory.Between1940and1982,theproduction of synthetic chemicals increased about 350 times. Billions of pounds ofsyntheticmaterialswerereleasedintotheenvironmentduringthisperiod.U.S.production of carbon-based synthetic chemicals topped 435 billion pounds in 1992,or 1600 pounds per capita (p. 137).6Theconceptofenvironmentalcontaminantsasestrogenicmimicsservestobring attention to the relationship between chemicals and ecological disruption. ThestructuralsimilaritybetweenDDTanddiethylstilbestrolisstriking.Theformerchemicalsubstancewasreleasedintotheenvironmentdecadesago,whereasthelatter was synthesized and marketed to pregnant women during the 1950s and thenusedasagrowthpromoterinlivestockuntilitwasbannedbytheFoodandDrugAdministration (FDA) in 1979.7At levels typically found in the environment, hormone-disrupting chemicals donot kill cells or attack DNA. Their target is hormones, the chemical messengers thatmove about constantly within the bodys communication. They mug the messengersor impersonate them. They jam signals. They scramble messages. They sow disin-formation.Theywreakallmannerofhavoc.Becausemessagesorchestratemanycriticalaspectsofdevelopment,fromsexualdifferentiationtobrainorganization,hormone-disrupting chemicals pose a particular hazard before birth and early in life(pp. 203204).6A more recent controversy has arisen around the apparent leaching of BisphenolAfromvarioussourcesofplasticsthatareinwidespreaduseamongconsumers.Earlier, the isolation and recovery of Bisphenol A from human urine was discussed.HowcouldthatmethodbechangedtoenableBisphenolAtobeisolatedandrecovered from an environmental matrix such as plastic wrap? Molecular structuresforp,p'-DDTanddiethylstilbestrolareshownbelow.ComparethesestructurestothatshownearlierinthischapterforBisphenolA.Thesimilaritiesinmolecularstructure are striking.The EPA has released its plan for testing 15,000 chemicals for their potential todisrupthormonesystemsinhumansandwildlife.Thesechemicalswerechosenbecause they are produced in volumes greater than 10,000 pounds per year.9p, p'-DDTCl ClClCl Cl OHOHDiethylstilbestrol 2006 by Taylor & Francis Group, LLC14 Trace Environmental Quantitative Analysis, Second EditionOne usually hears about environmental catastrophes through the vast resourcesof the mass media (i.e., radio, television, newspaper, popular magazines, newslettersfromspecialinterestorganizations,etc.).Themassmediausuallyassignsanametothedisasterthatalsoincludesageographicconnotation.ExamplesincludetheValdez Oil Spill in Alaska, Love Canal in New York, Seveso, Italy, and Times Beach,Missouri. Whatisnotsonewsworthy,yetmayhaveasprofoundanimpactontheenvironment, is the ever-so-subtle pollution of the environment day in and day out.Both catastrophic pollution and subtle pollution require the techniques of TEQA toobtaindatathatenablesocietytocontinuouslymonitortheenvironmenttoensureminimalecologicalandtoxicologicaldisruption.Itisthecombinationofsophisti-TEQA.This book provides insights and tools that enable an individual who either worksinanenvironmentaltestinglaborpublichealthlaboranticipateshavingacareerintheenvironmentalscienceorenvironmentalhealtheldtomakeacontribution.Individuals are thus empowered and can begin to deal with the problems of moni-toring and sometimes nding the extent to which chemicals have contaminated theenvironment or entered the human body.6. WHO NEEDS ENVIRONMENTAL TESTING?Itistooeasytoanswerthisquestionwitheveryone. TheindustrialsectoroftheU.S. economy is responsible for the majority of chemical contamination released totheenvironment.Sincetheearly1970s,industryhasbeenunderstateandfederalregulatory pressures not to exceed certain maximum contaminant levels (MCLs) fora variety of so-called priority pollutant organic and inorganic chemical substances.However, one of the more poignant examples of small-time pollution is that of drycleaningestablishmentslocatedinvariousshoppingplazasthroughouttheU.S.Thesesmallbusinesseswouldfollowthepracticeofdumpingtheirdrycleaninguid into their septic systems. It was not unusual, particularly during the 1980s, forlabstoanalyzedrinkingwatersamplesdrawnfromanaquiferthatservedtheshoppingplazaandndpartsperbillion(ppb)concentrationlevelsofchlorinatedvolatile organics such as perchloroethylene (PCE).ThenecessarysamplepreparationneededtomodifyasampletakenfromanaquiferthatisexpectedtocontainPCE,soastoenablethesampletobecomecompatible with the appropriate analytical instrument, will be described in Chapter 3.The identication and quantitative determination of priority pollutants like PCE indrinkingwaterrequiresophisticatedanalyticalinstrumentation.Theseso-calleddeterminative techniques will be described in Chapter 4. A laboratory exercise thatmightintroduceastudenttothetechniqueinvolvedinsamplepreparationandinstrumental analysis to quantitatively determine the presence or absence of a chlo-rinated volatile organic like PCE will be described in Chapter 5. 2006 by Taylor & Francis Group, LLCcatedanalyticalinstruments(Chapter4),samplepreparationschemes(Chapter3),mathematical treatment of analytical data (Chapter 2), and detailed practical proce-dures (Chapter 5) that enables a student or practicing analyst to effectively conductIntroduction to Trace Environmental Quantitative Analysis (TEQA) 157. WHO REQUIRES INDUSTRY TO PERFORM TEQA?Demandfortraceenvironmentalanalysisislargelyregulatorydriven,withtheexceptionoftheresearchdoneinmethodsdevelopmentbyboththeprivatesectorand federal, state, and academic labs. The major motivation for a company to conductTEQA is to demonstrate that its plants efuent falls within the MCLs for the kindsof chemical contaminants that are released. There exists a myriad of laws that governdischarges, and these laws also specify MCLs for targeted chemical contaminants.The following outline is a brief overview of the regulations, and it incorporates theabbreviations used by practitioners in this broad category of environmental compli-ance and monitoring (pp. 132).108. HOW DOES ONE MAKE SENSE OF ALL THE REGS?Thefollowingoutlinesummarizesthefederalregulationsresponsibleforenviron-mental compliance:A. Title40CodeofFederalRegulations(40CFR):Thisistheultimateauthorityforenvironmentalcompliance.Neweditionsof40CFRarepublishedannuallyandareavailableontheWorldWideWeb.Thisresourceincludeschaptersonair,water,pesticides,radiationprotection,noise abatement, ocean dumping, and solid wastes. Superfund, EmergencyPlanningandRight-to-Know,efuentguidelinesandstandards,energypolicy, and toxic substances are among other topics.B. GovernmentregulationsadministeredbytheEnvironmentalProtectionAgency (EPA)1. ResourceConservationandRecoveryAct(RCRA):PassageoftheRCRA in 1976 gave the EPA authority to oversee waste disposal andhazardouswastemanagement.Identicationofawasteashazardousreliedonspecicanalyticaltests.AnalyticalmethodsthatdealwithRCRAarefoundinacollectionoffourvolumestitledTestMethodsforEvaluatingSolidWastePhysical/ChemicalMethods,commonlyreferred to as SW-846. Three major subtitles deal with hazardous wastemanagement, solid waste management, and underground storage tanks.2. Comprehensive Environmental Response, Compensation, and LiabilityAct (Superfund) (CERCLA): This authority granted to the EPA enablesthe agency to take short-term or emergency action to address hazardoussituationsthataffecthealth.Thereleaseofthetoxicchemicalisocy-anate in the Bhopal, India, community that left over 3000 dead mighthave fallen under CERCLA if it had occurred in the U.S. In addition,the CERCLA contains the authority to force the cleanup of hazardouswaste sites that have been identied based on environmental analyticalresultsandplacedontheNationalPriorityList.TheEPAalsohasauthority to investigate the origins of waste found at these sites and toforcethegeneratorsandotherresponsiblepartiestopayunderCER-CLA. Analytical methods that deal with CERCLA are provided 2006 by Taylor & Francis Group, LLC16 Trace Environmental Quantitative Analysis, Second Editionthrough the Contract Laboratory Program (CLP). The actual methodsarefoundinvariousStatementsof Work(SOWs)thataredistributedto qualied laboratories.3. DrinkingWaterandWastewater:TheSafeDrinkingWaterAct,lastamendedin1986,givesEPAtheauthoritytoregulatedrinkingwaterquality.Twotypesofchemicalcompounds,calledtargetedanalytes,areconsideredinthisact. TherstistheNationalPrimaryDrinkingWater Standards. These chemical substances affect human health, andalldrinkingwatersystemsarerequiredtoreducetheirpresencetobelow the MCL set for each compound by the federal government. ThesecondistheNationalSecondaryDrinkingWaterStandards.Theseanalytes include chemical substances that affect the taste, odor, color,and other non-health-related qualities of water. A given chemical com-pound may appear on both lists at different levels of action. The primarytheMCL. Thiswasnotalwaystruehistoricallyintheeldof TEQAbecause available technology always serves to limit the MDL, whereasecological and toxicological considerations govern the decision to esti-mate what makes for an environmentally acceptable MCL.TABLE 1.3Primary Drinking Water Monitoring Requirements for InorganicsContaminant MCL (mg/L) MDL (mg/L)Antimony 0.006 0.00080.003Arsenic 0.05Barium 2 0.0010.1Beryllium 0.004 0.000020.0003Cadmium 0.005 0.00010.001Chromium 0.1 0.0010.007Copper 1.3 0.0010.05Cyanide 0.2 0.0050.02Fluoride 4Lead 0.015 0.001Mercury 0.002 0.0002Nickel 1.1 0.0060.005Nitrate-N 10 0.011Nitrite-N 1 0.0040.05Selenium 0.05 0.002Sodium 20Thallium 0.002 0.00070.001 2006 by Taylor & Francis Group, LLC1.5bycategory.Thesecondarydrinkingwatermonitoringcontami-drinking water monitoring contaminants are listed in Table 1.3 to Tabledetection limit (MDL). Note that the MDL should always be less thannants are listed in Table 1.6. The MCL is listed as well as the methodIntroduction to Trace Environmental Quantitative Analysis (TEQA) 17The Clean Water Act, which was last amended in 1987, provides forgrants to municipalities to build and upgrade treatment facilities. TheactalsoestablishesapermitsystemknownastheNationalPollutantDischargeandEliminationSystem(NPDES)fordischargeofnaturalwaterbodiesbyindustryandmunicipalities.Overtwothirdsofthestateshaveacceptedresponsibilityforadministrationoftheact.Theact and its amendments are based on the fact that no one has the rightto pollute the navigable waters of the U.S. Permits limit the compositionand concentration of pollutants in the discharge. Wastewater efuentsare monitored through the NPDES, and this analytical testing has beenTABLE 1.4Primary Drinking Water Monitoring Requirements for Semivolatile OrganicsContaminant MCL (mg/L) MDL (mg/L)Adipates 0.4 0.006Alachlor 0.002 0.002Atrazine 0.003 0.001Carbofuran 0.04 0.009Chlordane 0.002 0.0002Dalapon 0.2 0.001Dibromochloropropane 0.0002 0.000022,4-dichlorophenoxy-acetic acid 0.07 0.001Dinoseb 0.007 0.0002Diquat 0.02 0.0004Endothall 0.1 0.009Endrin 0.002 0.00001Ethylene dibromide 0.0005 0.00001Glyphosate 0.7 0.006Heptachlor 0.0004 0.0004Hexachlorobenzene 0.001 0.001Hexachlorocyclopentadiene 0.05 0.0001Lindane 0.0002 0.00002Methoxychlor 0.04 0.0001Oxamyl 0.2 0.002PAHs 0.0002 0.00002PCBs 0.0005 0.0001Pentachlorophenol 0.001 0.00004Phthalates 0.006 0.0006Picloram 0.5 0.0001Simazine 0.004 0.00007Toxaphene 0.003 0.0012,3,7,8-TCDD (dioxin) 0.00000003 0.0000000052,4,5-TP (Silvex) 0.05 0.0002Trihalomethanes (total) 0.1 0.0005 2006 by Taylor & Francis Group, LLC18 Trace Environmental Quantitative Analysis, Second Editionaboonforcommercialanalyticallaboratories.Everyindustryandwastewatertreatmentfacilitythatdirectsintoareceivingstreamorriver has either a federal or state NPDES permit. Methods of analysisare given in 40 CFR 136 for the following:(1) Conventional pollutants such as biological oxygen demand (BOD),chemicaloxygendemand(COD),pH,totalsuspendedsolids,oiland grease, and fecal coliforms(2) Nonconventionalpollutantssuchasnitrogen,phosphorous,andammonia(3) The 129 so-called priority pollutants4. Federal Insecticide, Fungicide, and Rodenticide Act (FIFRA): This actrequirestheEPAtooverseethemanufactureanduseofchemicalsubstances that directly terminate insects, fungi, and rodents. Analyticalmethods are required for the analysis of residues in air, soil, and water.Specications for labeling and warnings are also required under this act.5. Food,Drug,andCosmetic Act: AdministeredbytheFoodandDrugAdministration (FDA), this act governs all chemicals added to foods,drugs,andcosmetics.ThesesubstancesarenotdirectlyunderEPATABLE 1.5Primary Drinking Water Monitoring Requirements for Volatile OrganicsContaminant MCL (mg/L) MDL (mg/L)Benzene 0.005 0.0005Carbon tetrachloride 0.005 0.0005Chlorobenzene 0.1 0.0005p-Dichlorobenzene 0.075 0.0005o-Dichlorobenzene 0.6 0.00051,2-Dichloroethane 0.005 0.00051,1-Dichloroethylene 0.007 0.0005Cis-1,2-dichlorethylene 0.07 0.0005Trans-1,2-dichloroethylene 0.1 0.0005Dichloromethane 0.005 0.00051,2-Dichloropropane 0.005 0.0005Ethylbenzene 0.7 0.0005Styrene 0.1 0.0005Tetrachloroethylene (PCE) 0.005 0.0005Toluene 1 0.00051,2,4-Trichlorobenzene 0.005 0.00051,1,1-Trichloroethane 0.2 0.00051,1,2-Trichloroethane 0.2 0.0005Trichloroethylene (TCE) 0.005 0.0005Vinyl chloride 0.002 0.0005Total xylene 10 0.0005 2006 by Taylor & Francis Group, LLCThese pollutants are listed in Table 1.3 to Table 1.5.Introduction to Trace Environmental Quantitative Analysis (TEQA) 19oversight;however,thewastesandby-productsgeneratedfromthemanufacture of these substances are controlled by EPA.6. Toxic Substance Control Act (TSCA): Since 1976, the EPA has beengiven the authority to gather information on the toxicity and hazardousnatureofindividualchemicals.Chemicalproducersarerequiredtosupply information dealing with risk assessment of proposed products90 days before proposed manufacture or import. Included in this infor-mationarechemicalfatetesting,environmentaleffectstesting,andhealth effects testing.7. Superfund AmendmentsandReauthorization Act(SARA):Passedin1986, this act extends the lifetime of the legislation begun in CERCLAandgivestheEPAtheauthoritytoremediateasiteifnoresponsibleparties can be found to pay.8. Clean Air Act (CAA): The CAA amendments passed in 1990 gave theEPA authority to regulate many hazardous air pollutants (HAPs). Over100 of these HAPs must be regulated; they include well-known organiccompoundssuchasacetaldehyde,benzene,carbontetrachloride,1,4-dioxane,hexane,methylmethacrylate,andsoforth.TheCAAalsorequires the EPA to establish permits for the regulation of the maximumamounts of emissions by various industries in a manner similar to thatof the NPDES program for wastewater efuents.Asifthenumberofregulationsisnotenough,manyenvironmentaltestinglabsalso have to satisfy state requirements. Most states run their own private lab certicationTABLE 1.6Secondary Drinking Water Monitoring RequirementsContaminant MCL (mg/L)Aluminum 0.050.2Chloride 250Color 15 color unitsCopper 1.0Corrosivity NoncorrosiveFluoride 2.0Foaming agents 0.5Iron 0.3Manganese 0.05Odor 3 TONpH 6.58.5Silver 0.1Sulfate 250Total dissolved solids (TDSs) 500Zinc 5 2006 by Taylor & Francis Group, LLC20 Trace Environmental Quantitative Analysis, Second Editionprograms. Also,someofthemethoddetailsdifferbetweenstateandfederalpro-grams. One example from the authors own experience working in an EPA contractlabduringthelate1980sinvolvestheimplementationofEPAMethod8270forsemivolatile organics in solid waste. EPA Method 3640A, Gel Permeation Chroma-tography (GPC), is added to any 1 of the 3500 series of sample preparation methods,particularly if soil samples are saturated with oil. To satisfy the New Jersey Depart-mentofEnvironmentalProtectionrequirementswhenthismethodisimplementedrequires that the GPC cleanup step not be used even if some soil samples that arrivetothelaboratoryareindeedsaturatedwithoil.Ontheotherhand,ifsimilaroil-ladensoilsampleswereanalyzedfortheNew YorkDepartmentofEnvironmentalConservation, GPC cleanup is acceptable. What is an analyst to do?9. WHAT ANALYTICAL METHODS SATISFY THESE REGULATIONS?Today,myriadsofanalyticalmethodsexistthattheabove-citedregulationsusetothe MCL and MDL, are only obtained with good accuracy and precision by applyingall of the skills and techniques of TEQA. The analyst who is about to work in theenvironmental testing laboratory must have access to these written methods in ordertobecomefamiliarwiththeircontentbeforeheorshecaneffectivelyexecutethemethodsthemselves.TheEPAduringthepast30yearshasoperatedwithintwobroad mental frameworks or paradigms. The rst paradigm, which began in the earlyTABLE 1.7Method Series 00009999: Test Methods for Evaluating Solid Wastes Physical/Chemical MethodsMethod Description0000s Air emission sampling methods from stationary sources1000s Methods to determine hazardous waste characteristics: ignitibility, corrosivity, and reactivity; includes the toxicity characteristic leaching procedure (TCLP)2000s Unused3000s Sample preparation methods for inorganics and organics4000s Soil screening methods by immunoassay5000s Sample preparation methods for volatile organics and miscellaneous sample preparation methods6000s Methods to determine more than one metal at a time7000s Methods to determine one metal at a time8000s Methods to determine organics9000s Miscellaneous test methods; includes cyanides, suldes, phenols, oil and grease, chlorine, total coliform, potentiometric halides and cyanide, colorimetric chloride and radium-228 isotopeSource:TestMethodsforEvaluatingSolidWastes,Physical/ChemicalMethods,SW-846,3rded.,Supplement III Update, July 1997. 2006 by Taylor & Francis Group, LLCdemonstrate compliance. The two parameters cited in Table 1.3 to Table 1.5, namely,Introduction to Trace Environmental Quantitative Analysis (TEQA) 211970sandlasteduntiltheearly1990s,suggestedthatthemethodswrittenbytheagency were the only ones that a laboratory could use in order to satisfy the regs.This mind-set led to the proverbial EPA approved stamp of approval. Instrumentmanufacturers during the late 1980s would incorporate into their advertisements thattheirproductwasEPAapproved.Duringtheseyears,instrumentvendorssoughtcollaborationswithvariousEPAofcesinanattempttoacceleratethemorethan10-year delay between proposal of a new method and its acceptance by the EPA.For example, the development of an instrumental method that would determinemore than one inorganic anion in a drinking water sample was announced in 1975.11It took until around 1985 for this technique to become incorporated into EPA Method300.0 as applied to drinking water. Prior to the development of ion chromatography,individualanionicspeciessuchaschloride,nitrate,phosphate,andsulfateweredetermined individually by applying specic colorimetric methods. Each colorimet-ricmethodwouldrequireseparatesamplepreparationanddeterminativestepsculminatingwiththeuseofaspectrophotometer. Theideathatasamplecouldbeinjectedintosomekindofchromatographandalloftheionsseparated,detected,and quantitated in a reasonable period represented a bit of a revolutionary concept.Itwasalsothoughtthattherewouldbeaseriousmatrixeffectthatwouldrenderthis new method useless when it came time to analyze dirty samples. Today, aftersome 20 or more years of instrumentation renement, the advantages and limitationsof the method that requires ion chromatography are well understood.11 This techniquehasevenadvancedtothepointwhereonecanconductanalyseswithouthavingtoprepare either an eluent (mobile phase passed through the ion exchange column) ora regenerate (mobile phase passed through the suppressor), thus further simplifyingthe implementation of this methodology.12ShouldittakeadecadeortwoforanewlydevelopedmethodtobecomeEPAapproved?Thefutureappearstonotbeemulatingthepast,asthesecondbroadmental framework is taking hold; the agency calls it the performance-based method(PBM).Thenewparadigmplacesmoreoftheresponsibilityforqualityassuranceonthe individual analyst and the laboratory rather than on the method itself. It used tobe that as long as an analyst followed the recipes (sounds a lot like cooking) in thevarious methods, then quality data were assured. The analysts input in the processwasnotrequired,norwasitrequested.Thispointofviewtooktheanalystoutofthepictureandservedtodenetheanalystmoreasarobot-technicianthanasaninvolvedandmotivatedprofessional.Thispointofviewleftnoroomforanalystintervention. If an analyst wished to skip a step here or there in the method, it wasassumedthatthemethodwasnotfollowedand,therefore,thedatacouldnotbeassuredor,intheparlanceofthetime,EPAapproved.PBMshopefullyhaveandwill change this somewhat myopic view of TEQA and, furthermore, begin to placetheresponsibilityontheanalysttoyieldqualitydatathatareassuredbasedonmethod performance, as opposed to blindly adopting the recipe.Forexample,insteadofanEPA-approvedmethodspecifyingthataparticulargaschromatographic(GC)columnbeusedtomeasureanalyte AinmatrixB,thelaboratory is free to choose what column to use, as long as an equivalent degree ofquality assurance is obtained. In other words, analytical results for the concentration 2006 by Taylor & Francis Group, LLC22 Trace Environmental Quantitative Analysis, Second Editionof analyte A in matrix B from the approved method vs. the PBM approach shouldbe nearly identical. In addition, the laboratory would not have to conduct the side-by-sidecomparisonofamoreconventionalsamplepreparationagainstaproposedalternative in this new paradigm. For example, it was once considered heresy for ananalysttoanalyzewastewatersamplestodeterminetheslewoforganochlorinepesticides using anything except a glass column that contained Supelcoport coatedwith 1.5% SP-2250/1.95% SP-2401 packed into a 1.8-m-long 4-mm-inner-diametertube (the essence of Method 608). However, during the late 1980s, labs, includingthe one this author worked in, began to investigate megabore capillary columns asalternatives to packed columns. Megabore capillary columns made from fused silicacouldbeeasilyconnectedtothecommon-in.injectionportusedforpackedcolumns via megabore adapter kits. When combined with element-specic detectors,many new organics could be separated and identied using megabore capillary GCcolumns.13Performance-based methods are an empowering concept. PBMs enable analystsandthelaboratoriesthattheyworkintobecomemorecreativeandencourageattempts to develop alternative methods to perform TEQA. Many of these alternativeapproaches are much more cost effective while yielding the same outcome. In otherwords,theperformanceofthemethodisthesame,deliverableoutcome,andtheinputs are much reduced in cost and labor. Alternative methods of sample preparation10. HOW ARE THE REGULATORY METHODS FOR TEQA IDENTIFIED?AllEPAmethodsareclassiedbasedonanumber,whereasothermethodsarecategorized based on the chemical composition of the analytes of interest. It is usefulatthispointtobrieyoutlinetheseEPAmethodnumberswhileprovidingabriefdescriptionoftheirorigin(pp.3336).10AllofthemajorofceswithintheEPAhave had a role in developing analytical methods that bear a number. Series 1 to 28digits refer to methods for the chemical analysis of water and wastes and representsomeoftheearliestEPAmethods.The100seriesofmethodscharacterizethethe 300 series deals with inorganics and nonmetals. This is why the ion chromato-graphic technique discussed earlier is found in this series. The 400 series deals withorganics where the emphasis is on nonspecic organics that are primarily found inwastewater.Method413foroilandgrease,Method415fortotalorganiccarbon,Method 420 for total recoverable phenols, and Method 425 for methylene blue activesurfactants are examples of nonspecic analytical methods that give good indicationsof whether a waste sample contains organics without the need to be specic. Theseare workhorse methods that are usually found in most environmental testing labs.Method413hasrecentlybecomecontroversialinthatthemanufactureofthecommonextractionsolventusedinthemethod,1,1,2-trichlorotriuoroethane 2006 by Taylor & Francis Group, LLCwill be discussed in Chapter 3.areairmonitoringmethodsfoundin40CFR Appendix A,whereasseries101to115areairmonitoringmethodsin40CFR AppendixB.Methodsthathavethreephysicalpropertiesofwaterandwastes.The200seriesisdevotedtometals,andIntroduction to Trace Environmental Quantitative Analysis (TEQA) 23(TCTFE), itself a uorocarbon (Freon 113), is banned. The attributes of TCTFE aresuch that it enabled this method to move from a purely gravimetric approach (EPAMethod413.1,Gravimetric,SeparatoryFunnelExtraction)toaninstrumentalapproach,wherebythecarbon-to-hydrogenstretchingfrequencyintheinfraredspectrum enabled a signicant reduction in the MDL to occur. The infrared approachwasassigneditsownmethodnumber,EPAMethod413.2,Spectrophotometric,Infrared.ThisauthorspersonalopinionisthatthemorecosteffectiveinfraredapproachcoupledwiththelowerMDLsshouldbemaintained.Labsshouldbestrongly encouraged to recycle the waste TCTFE solution so that it can be reused.AsimpledistillationshouldsufcetoreturnTCTFEinsufcientpuritytousetoreextractsamples.However,theEPAhaschosentoestablishMethod1664foroiland grease analysis. TCTFE has been replaced by hexane, and the determination isby weighing the residue after solvent evaporation.Onegoodthingaboutthisnewmethodisthatitisperformancebased.Theanalyst can use an alternative approach to oil and grease isolation and recovery. Oneapproach is to use solid-phase extraction (SPE) to isolate the oil and grease, whereastheotherapproachistondawaytoeliminateuseofthesolvent.Therecentlyintroduceduseofinfraredcardsby3MCorporation,wherebytheoilandgreaseremainonathinlmofTeon,isanexampleofthelatteralternativetotheconventionalliquidliquidextraction(LLE).Thethinlmthatnowoffersaxedpath length is then inserted into the sample holder of a conventional infrared spec-trophotometer, and the absorbance of the CH stretching vibration at 2900 cm1 isdirectlyrelatedtoconcentrationviatheBeerLambertlawofspectrophotometry,The 500 series methods refer to measurement of trace organics in drinking water.This is a collection of methods rst published in the late 1980s and incorporating anumberofthemoreinnovativetechniquesintroducedatthattime.The600seriesmethods describe how to isolate, recover, and measure trace organics in wastewaters.These methods were rst promulgated in the 1970s. Both series of methods measurespecic organic compounds and require a means to separate, identify, and quantifytheseorganics. Thus,samplesthatmaycontaintwoormorecompoundsrequireachromatographicseparation.Hence,mostallmethodsinthesetwoseriesareGCmethods. Thescopeofbothmethodsisalsolimitedtoorganicsthatareamenableto GC. Sample preparation for trace volatile organics (commonly abbreviated VOCs)Chapter 4.The 900 series of methods refer to the measurement of radioactivity in drinkingwater. These methods were rst promulgated in 1980.The four-digit series, methods 0000 to 9999, involve the analysis of solid wastes.Thisserieswaspromulgatedintheearly1980sandhasundergoneseveralmajorrevisions up to the present. This series, when compared to all of the others, seemstobethemostdynamicandisconsideredbytheEPAasageneralguide.Theseviewpoints infer that these methods were written within the framework of the secondparadigm. In other words, the methods themselves are performance based. The seriesisperiodicallyupdatedandrevised,andnewmethodsarecontinuouslyadded.AnovicetotheeldofTEQAwoulddowelltoinitiallyfocusonthisseriesasa 2006 by Taylor & Francis Group, LLCdiscussed in Chapter 4.willbeintroducedinChapter3,andtheprinciplesofGCwillbeintroducedin24 Trace Environmental Quantitative Analysis, Second Editionlearningtool.Atableofcontentsforthisseriesofanalyticalmethods,whichis14 To illustrate thedynamic nature of the SW-846 series of analytical methods for TEQA, consider thevery recent development of Method 7473 for determining mercury in various solidsof environmental interest.15 A solid sample such as apple leaves, oyster tissue, coalyash,orriversedimentisdriedandthermallydecomposedat750Cinanoxy-genatedfurnace.Mercuryvaporisreleased,sweptthrough,andtrappedonagoldamalgamator. Theamalgamisthermallydesorbed,andtheHgvaporissweptintoa cold vapor atomic absorption spectrophotometer, where the absorbance is measuedat254nm. Again,theBeerLambertlawisusedtorelateHgabsorbancetocon-centration. An instrument detection limit (IDL) is reported to be 0.01 ng of total Hg.One of the most challenging methods to implement in the environmental testinglaboratoryisEPAMethod8270.Thisisamethodthatseparates,identies,andquanties over 100 priority pollutant semivolatile and nonvolatile organics in varioussolidwastes.Themethodisadeterminativeoneandisusedincombinationwithone of the sample preparation methods found in the 3000 series of SW-846. A briefdiscussion of this method follows in order to introduce the reader to the challengesofthismethod,identiedas8270C. WhenMethod8270wasrstpromulgated,anumberofanalytesfromthetargetedlistfoundinthemethodeitherdidnotyieldahighrecoveryordidnotchromatographwell.Thisresultedintheanalystbeingunabletosatisfythecriteriaandledtoarejectionoftheanalyticalresultsinthevalidationprocess.Method8270Cclearlystatesattheonsetthatcertainanalytesarenondetectedbythismethodorhaveanunfavorabledistributioncoefcient(toduringsamplepreparation.ThethreerevisionsofthismethodservetoillustratenicelyhowtheEPAhasbeenabletoreadjustitspromulgatedmethodsunderthenew PBM paradigm to address early aws. Under the old paradigm, this realizationon the part of the EPA was not evident.The method itself is succinctly summarized as follows:Method 8270 can be used to quantitate most neutral, acidic or basic organic compoundsthat are soluble in methylene chloride and capable of being eluted, without derivatiza-tion, as sharp peaks from a gas chromatographic fused-silica capillary column coatedwithaslightlypolarsilicone.Suchcompoundsincludepolynucleararomatichydro-carbons,chlorinatedhydrocarbonsandpesticides,phthalateesters,organophosphateesters, nitrosamines, haloethers, aldehydes, ethers, ketones, anilines, pyridines, quino-lines, aromatic nitro compounds and phenols including nitrophenols.1611. WHY IS IT CONSIDERED A CHALLENGE TO IMPLEMENT AN EPA METHOD?The most tedious and time consuming aspects of Method 8270 are involved in thepreparationandorganizationofthevariousreferencedchemicalstandards. Thisisin addition to the use of reference standards that are required in the various sample 2006 by Taylor & Francis Group, LLCof implementing such a method. The most current SW-846 series lists a third revisioncollectively known as SW-846, 3rd edition, is given in Table 1.7.bediscussedinChapter3)oradsorbtotheinnerwallsofglasswareorhydrolyzeIntroduction to Trace Environmental Quantitative Analysis (TEQA) 25preparation methods (i.e., the 3000 series). Method 8270C requires that the followingcategoriesofreferencestandardsbeprepared,maintained,andreplenished,whennecessary, by the analyst:1. Stock reference standards prepared in the laboratory from the dissolutionof neat forms of the individual targeted chemical compounds or purchasedfrom various suppliers. The EPA used to provide these from its repositorylocated in Research Triangle Park, NC, but no longer does.2. Internalstandardreferencesolutionsthatcontain,withtheexceptionof1,4-dichlorobenzene-d8, ve deuterated polycyclic aromatic hydrocarbonsmustbeprepared.Table1.8,adaptedfromthemethod,showswhichofthesemivolatileorganicsaretobequantitatedagainstphenanthrene-d10as the internal standard. The analytes listed in Table 1.8 all elute from thecapillary column over a range of retention times that are close to that ofphenanthrene-d10.3. GC-MStuningstandardforsemivolatilesthatcontainsthetuningcom-pounddecauorotriphenylphosphine(DFTPP)andalso4,4-DDT,pentachlorophenol, and benzidine. DFTPP is used to verify that the hard-ware tune was carried out previously, and the other three analytes are usedtoverifyinjectionportinertnessandGCcolumnperformance.Polarcompounds like phenols tend to adsorb to contaminated surfaces and resultin the complete loss of response or poor peak shape.4. Dilutionofstocksolutionstopreparetheseriesofworkingcalibrationstandards that are injected into the gas chromatograph-mass spectrometer(GC-MS)tocalibratetheinstrumentforeachofthe100ormoresemi-volatile organics. This series of standards should also contain at least oneinitialcalibrationverication(ICV)standardtoassesstheaccuracyandprecision of the calibration curve.5. Surrogate standard solutions whose analytes are chemically different fromany of the targeted analytes and are added to each sample to assess percentrecovery.TABLE 1.8List of Priority Pollutant Semivolatile Organics Quantitated with Respect to the Internal Standard Phenanthrene-d104-Aminobiphenyl HexachlorobenzeneAnthracene N-Nitrosodiphenylamine4-Bromophenyl phenyl ether PentachlorophenolDi-n-butyl phthalate Pentachloronitrobenzene4,6-Dinitro-2-methyl phenol PhenancetinDiphenylamine PhenanthreneFluoranthene Pronamide 2006 by Taylor & Francis Group, LLC26 Trace Environmental Quantitative Analysis, Second Edition6. Matrix spike standard solutions and laboratory control standards that areadded to one sample per batch to assess matrix effects.7. System performance check standards and calibration check standards areused to ensure that minimum response factors are met before the calibra-tion curve is used.8. Blanksthatrepresentstandardsthatcontainnotargetedanalytes,surro-gates,ormatrixspikesandareusedtoassessthedegreetowhichalaboratory is contaminated.All of these standards must be available to the analyst in the laboratory. A mostThe requirement that the analytes be quantitated using an internal standard modeof instrument calibration is due to the fact that mass spectrometers are intrinsicallyunstable; that is, their response factor varies with time when compared to other GCdetectors,suchastheameionizationdetector(FID).Theinternalstandardtech-12. WHAT MADE EPA METHOD 625 SO UNIQUE?EPA Method 625 discusses exactly how to isolate, recover, and quantitate the variouspriority pollutant S VOCs from a wastewater matrix. The 600 series of EPA methodsincludebothsampleprepanddeterminativetechniquesincontrasttotheSW-846methods.Itwasrealizedbackintheearly1970sthatprioritypollutantanalyteidentication (answers the trace qualitative analysis question) based only on analyteretention time (the time it takes for a given analyte or chemical compound to passthrough the chromatographic column after being injected; retention time is usuallygiventhesymboltR)wasinadequateforregulatorypurposesrelyingonlyongaschromatographs with element-specic or so-called standard detectors. According toBudde (p. 171):17The proposal of Method 625 for regulatory use was very signicant and had a majorimpact on the subsequent practice of environmental analyses and the development ofcommercial GC-MS instrumentation. The prospect of using a single analytical methodwithnorequiredextractfractionationfor83analyteswasattractiveeventhoughitrequiredamorecostlyanalyticalinstrument. Thealternativewastheimplementationof eight different GC columns and detectors. The economic implications were clear tothe regulated industries and the emerging environmental testing industry.and recover the original 83 priority pollutants from wastewater. Prior to the devel-opmentofcapillaryGCcolumns,theuseofpackedGCcolumnsnecessitatedthattwo fractions be prepared. A base-neutral fraction was prepared and injected into aGC-MS,followedbyanacidfraction. TheorganicphasewasfurtherevaluatedasusesaGCincorporatingaameionizationdetector(GC-FID).TheFIDwillbedescribed in detail in Chapter 4, sufce to say here that this detector is universally 2006 by Taylor & Francis Group, LLCScheme 1.2 introduces the sample preparation rationale that was used to isolateshown in Scheme 1.3. Based on how colored the extract is, a preliminary screennique of instrument calibration is discussed in Chapter 2, and the principles of massimportant aspect of an analysts job is to keep track of all of these standards.spectrometry are introduced in Chapter 4.Introduction to Trace Environmental Quantitative Analysis (TEQA) 27selective for organic compounds whose molecules contain a carbonhydrogen cova-lent bond. The aqueous phase from Scheme 1.2 that contains the conjugate base forthe organic acids, such as phenols, is acidied, extracted, and also evaluated basedthroughoutthisbooktobrieydescribesamplepreparationtechniques. Theirpur-pose is to help facilitate a quicker understanding of the many approaches to sampleSCHEME 1.2SVOCs (BNAs) in wastewateraccording to EPA Method 625 Is an emulsion expected? No Adjust sample pH to >11;add surrogates; add BNA matrix spikeAdd appropriate extracting solvent or mixed solvents Extract twice more and combine extracts No Yes Transfer sample or extract to a continuous extractor apparatus Has an emulsion formed? Yes Organic Phase: so-called base-neutral containing phase

Aqueous Phase (save!):so-called organic acids containing phase 2006 by Taylor & Francis Group, LLConthecoloroftheextract,asshowninScheme1.4.Flowchartswillbeused28 Trace Environmental Quantitative Analysis, Second Editionpreparation. Budde summarizes and further comments on Method 625 from a con-temporary perspective, as follows (p. 173):17Method625waspromulgatedinwastewaterregulationson26October1984.ThemethodrequiresthepHadjustmentsbeforeextractionandspeciestwopackedGCcolumns. However, the method does allow the use of capillary GC columns, but doesnotprovideinformationtoassisttheanalystintheapplicationofthesecolumns. Asof1999,Method625hasnotbeenrevisedtomodifythepHadjustmentsbeforeextraction,provideinformationaboutfusedsilicacapillarycolumns,orincorporateother improvements in analytical techniques.SCHEME 1.3Is extract colored? No Inject into GC-FID, interpret chromatogram No peaks or peaks < 100% of full-scale deection Calculate a dilution factor Peaks > 100% of full-scale deection Proceed to analyze undiluted extract by GC-MS assuming DFTPP criteria has been metYes Reduce extract volume via Kuderna- Danish evaporative concentrator Remove residual water Organic phase containing base-neutrals SVOCsaccording to EPA Method 625Proceed to analyze diluted extract by GC-MS assuming DFTPP tuning criteria has been met 2006 by Taylor & Francis Group, LLCIntroduction to Trace Environmental Quantitative Analysis (TEQA) 2913. WHAT ABOUT METHODS FOR THE ANALYSIS OF AIR AND OTHER METHODS?This book clearly focuses on sample matrices drawn from condensed states of matter.However, there is a set of established methods chiey to monitor targeted VOCs inmethodsalongwiththerequiredsamplinganddeterminativetechniques.Thesemethodsareconventionalairsamplingtechniques. Arecentandexcitingadditionto ambient air sampling is to use a solid-phase microextraction ber as a samplingSCHEME 1.4Aqueous phase containing acidic organics such asphenols according to EPA Method 625 Adjust pH of aqueous phase to