abstracts of papers presented at the 2002 pittsburgh...

Abstracts of papers presented at the 2002Pittsburgh Conference

The following 125 abstracts form Part B of two issues of Journal of Automated Methods & Management in Chemistrydevoted to abstracts of papers and posters presented at the Pittsburgh Conference held from 17 to 22 March 2002 inNew Orleans, LA, USA. The papers and posters covered a range of topics and techniques, each of which providedvaluable information to the conferees and exhibitors alike. The attendance ® gures remained disappointing, but Pittcon isstill one of my favourite shows and has done a great job for Analytical Chemistry as a whole. People are tiring of NewOrleans as a venue and it has become little more than a trade show. Hopefully, it will revive itself when the next showwill be held in Orlando, FL, from 9 to 14 March 2003. Perhaps the attendees will spare some time from Disney and SeaWorld to visit us at the exhibition and conference.

If you need further information about any of these abstracts, please contact the authors. It is my intention to publish fullpapers corresponding to some of these abstracts in future issues of the journal.

Strategies for successful global laboratory infor-mation management systems implementations

Stuart M. Miller, Taratec Development Corporation, LIMS &Lab Automation Solutions, 1170 US Highway 22, Ste 302,Bridgewater, NJ 08807-2933, USA

Pharmaceutical and life sciences companies world-wideare under enormous pressure to transform their drugdiscovery and development processes in order to makedecisions on whether to fail or promote compounddevelopment with more con® dence and in a muchshorter period of time. Computing and software areplaying a vital role in this transformation and as a® rst step, many life sciences companies are establishingfoundations of globally harmonized information systemsthat are intimately tied to the key business processes.LIMS are one of the key systems that provide muchof the data required to make the go/no go decisions,and are often one of the ® rst areas of focus for theseglobal harmonization initiatives. Successfully completinga truly global LIMS project involves the usual arrayof technical challenges but the regulated environmentsof the life sciences industry and the diverse make up ofthe global teams present additional challenges.Simultaneously satisfying FDA and internationalregulatory requirements with a uni® ed global systemand establishing common goals and harmonization ofthe underlying business processes which meet therequirements of the local sites are typically two ofthe greatest impediments to the success of theseprojects. While life sciences companies are eager toreap the bene® ts of a truly global LIMS, special caremust be taken to ensure the global teams are preparedto handle these and other challenges that often putthe success of these critical projects at risk of producingmerely another set of regionally diverse businessprocesses and information systems. In this presentation,we will recommend proven strategies to adopt andpitfalls to avoid in the planning and execution of aglobal LIMS implementation in the evolving lifesciences enterprise. Contrasting anonymous casestudies will be used to illustrate the dos and don’ ts ofthis process.

An LIMS to support recent developments in stabi-lity testing

Nick Townsend and Martin Waugh, Autoscribe, Ltd, 81 Tech-nology Park Drvie, East Falmouth, MA 02536, USA

Stability testing is becoming more important in gainingregulatory approval to market new formulations ofexisting drugs and new drug products. Some companieshave begun testing the stability of all drug intermedi-aries as well as the raw materials and the ® nal packagedproduct. With the acceptance of standardized storageconditions of temperature and humidity, by severalinternational regulatory agencies, there is a need toreplicate commonly used protocols in new studies.Furthermore, many protocols now include ` conditioncycling’ as part of the study, so as to simulate real-lifesituations better. These new requirements have beenaccompanied by new regulatory guidelines for electronicrecord-keeping (21 CFR Part 11) We will presentinformation on a new LIMS that has been designed incooperation with several pharmaceutical companies tomeet these new requirements. It uses an intuitivegraphical interface to design and approve new protocolsand studies. Container/closure, condition and pull li-braries are used, as are study and batch copy functions,to speed the set up and approval of new batches. Thesystem manages container inventories, including a provi-sion to track spare containers and scrap any remainingcontainers when a study is completed. Examples ofscreens and reports will be provided along with a fulldescription of how the system maintains an audit trail ofall activities.

Delivering laboratory information in an Internetsociety

Mark E. Fish1 and Hemal Rajani2, 1Thermo LabSystems, 1 StGeorge’s Court, Altrincham WA14 5TP, UK, 2Thermo Lab-Systems, Analytical Chemistry, Caixa Postal 6154, Campinas,Sao Paulo 13081-970, Brazil

The emergence of the Internet has been the mostsigni® cant computing development in recent times, ifnot ever. This has caused an explosion in the amount

Journal of Automated Methods & Management in ChemistryVol. 24, No. 6 (November–December 2002) pp. 171–208

Journal of Automated Methods & Management in Chemistry ISSN 1463± 9246 print/ISSN 1464± 5068 online # 2002 Taylor & Francis Ltdhttp://www.tandf.co.uk/journals

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of information available on a global scale. The transfer ofinformation and data is a fundamental aspect of ana-lytical sciences. Making this information easily availableis an essential requirement from bench to board leveland can oå er signi® cant competitive advantages in anincreasingly competitive market place. An intranet todaycan be used to create new interfaces for complex decisionsupport systems, drawing from a pool of many previouslyunconnected databases. In the world of laboratoryautomation, as with any type of industry, the oppor-tunity of enhanced productivity by improving the ¯ ow ofinformation and the development of cost-e å ective sol-utions for sharing company information are welcomedwith open arms. This paper will examine tools andtechnological available for companies to deliver labora-tory information in both Intranet and Internet environ-ments. The paper will also discuss the bene® ts andpotential pitfalls of delivering laboratory information inan Internet society.

Electronic laboratory notebooks and electronicrecord-keeping systems

Kevin Smith, Thermo LabSystems, R&D Department, 1 StGeorge’s Court, Hanover B, Altrincham WA14 5TP, UK

For hundreds of years scientists have relied on their papernotebooks as a medium for recording their work. Manyhave dreamed of a paperless solution, but the market forElectronic Lab Notebooks remains in its infancy. It isThermo LabSystems ’ view that the technology is nowavailable to enable development of a comprehensiveElectronic Lab Notebook and as a result we have beenperforming some market research involving end users,other vendors, and industry groups such as CENSA(http://www.censa.org) We have categorized the require-ments into three broad areas:

. Ease and convenience of use, including mobility,compound document creation, instrument integra-tion, etc.

. Record keeping: addressing regulatory (21 CFRPart 11) and IP issues arising from the long-termstorage of records in electronic format.

. Knowledge management: allowing scientists tocollaborate through publishing, sharing, andmining the above records, again over the long-term.

The ® rst set of these requirements is relatively easy tomeet and there are many current examples (web tablets,word processing packages, LIMS, etc.). However, long-term record keeping and knowledge management re-quire data to be stored in a back end system using` platform neutral ’ formats, allowing data to be readafter the original application is no longer available.For human readable data, such as text and diagrams,Adobe PDF is a widely accepted format, expected to bereadable for many years into the future. For analyticaldata generated by instruments such as GC/LC, MS, FT-IR, UV, NMR, etc., eXtensible Markup Language(XML) is more suited and again is likely to be readablein many years to come. Based on the above, we arecurrently working on an Electronic Record Manage-ment (ERM) system which we believe will be suitable

(1) as a backend for an ELN and (2) as a standalonesystem for long-term laboratory data archival. Thepresentation will describe the above issues and possiblesolutions in more detail.

Chemometric data evaluation for improved sea-water monitoring of chlorinated hydrocarbons bymid-infrared evanescent wave sensors

Frank Vogt1, Manfred Karlowatz1, Lubos Hvozdara1, BorisMizaiko¡1 and Martin Kraft2, 1Georgia Institute of Technology,School of Chemistry and Biochemistry, 770 State Street NW,Atlanta, GA 30332-0400, USA, 2Vienna University of Tech-nology, Institute of Analytical Chemistry, Getreidemarkt 9/151,Vienna A-1060, Austria

The increasing pollution of the world water resourcesand the marine ecosystems demands for novel on-lineand in-situ measurement techniques. Scienti® c goal is toenable concentration surveillance and enhanced under-standing of pollutants transport mechanisms. Based on aBruker Vector 22 mid-infrared FT-IR spectrometer, anovel sensing device is developed for pollutant detectionin seawater. This device is designed for submarinemeasurement tasks by installing a FT-IR spectrometerin a deep-sea housing, which is connected to a polymercoated silver-halide waveguide sensor. This sensor usesthe principle of mid-infrared evanescent wave sensing.At the time, the target analytes dissolved in seawater attrace concentrations are chlorinated hydrocarbons , likechlorobenzene, dichlorobenzene, tri- and tetrachloro-etheylen, as well as aromatic hydrocarbons includingbenzene, toluene and xylene isomers. Previous investiga-tions have already demonstrated the feasibility of thisset-up for measuring such compounds under real worldconditions. This paper is focused on the evaluation ofoverlapping spectral features of diå erent pollutantsusing chemometric algorithms, like principal componentregression (PCR), for improved concentration determi-nation in multicomponent seawater samples. The in u-ence of the salinity and the turbidity on the spectra isalso discussed. For the ® rst time, pollutants in marineecosystems can be determined on-line and in-situ withhigh precision, i.e. in the ppb concentration range. Thedevelopment of the ® rst mid-infrared sensing system forsubmarine applications signi® cantly enhances the ana-lytical methodology for marine monitoring and opensan entirely new ® eld for spectroscopic chemical sensorsystems.

A new UV on-line photometer for low level NOx

measurement from emission sources

Bill Worthington1 and Walter Fabinski2, 1ABB, Inc., Analy-tical Department, 843 N. Je¡erson Street, Lewisburg, WV24901-9509, USA, 2ABB, Analytical, Stierstaedter Str, Frank-furt am Main D-60488, Germany

Nitrogen oxides have a particular relevance as anenvironmental component with a global eå ect. Themain sources are combustion engines and stationarycombustion processes. The global reduction of NOx ismandated since progressing knowledge about the eå ectof nitrogen oxides on the climate and the biosphere are

Abstracts of papers presented at the 2002 Pittsburgh Conference

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proven. In this eå ort a further reduction of the emissionlimits for stationary sources is sought which leads tomeasuring ranges of µ10 ppm Knox. This new approachleads many users to look for alternative measuringtechniques to meet the current challenge of these lowerrange requirements. In addition, a robust and reliablemeasuring technology with long maintenance intervalsis always desired. This paper describes a new develop-ment of a new UV photometer for the measurement ofNOx in applications in CEM and in the automobileindustry. The technology is based on a UV-dischargelamp that delivers NO-speci ® c UV-radiation. Since theemitted radiation is in resonance with the NO absorp-tion lines, it results to high sensitivity and selectivity.This unique technique is called UV-RAS (ultravioletresonance spectrometer). A four-channel signal pro-cessing gives a high stability. Since the measurementdoes not need any vacuum pump, ozonizer/de-ozonizerand auxiliary gases, it is robust and reliable and needslittle maintenance. The analyser has the potential for ameasuring range of 5± 10 ppm NOx. We will reportabout the results of tests run in the automotive industryand on turbine exhaust gases.

HRMAS NMR as an analytical tool in combina-torial chemistry

Guy Lippens, R. Warrass, P. Rousselot-Pailley, J. Martins, G.Chessari and J.-M. Wieruszeski, Institut Pasteur de Lille,Institut de Biologie de Lille, NMR Laboratory, Lille F-59000,France

At the heart of the present day eå ort in drug discoveryand development are the modern methods for stereo-selective organic synthesis. A breakthrough has been theintroduction of combinatoria l chemistry, where largenumbers of compounds are synthesized in a relativelyshort time. Less based on fundamental principles butrather on practical considerations, one could draw adivision in this emerging ® eld between solution andsolid-phase methods [1]. One of the chemical advantagesof solid-phase organic chemistry (SPOC) is the possibilityof automation turning it into a method of choice forcombinatorial chemistry. However, transposing theknown solution reaction schemes to the solid-phase hasnot been straightforward in all cases. Therefore, it isnow generally recognized that optimizing the reactionconditions is the most time consuming step. One of thedisadvantages in this process is the lack of universal andrapid analytical methods for on-resin quantitative analy-sis of the solid-supported compounds, both at intermedi-ate stages and of the ® nal products, while avoiding thetedious and time-consuming procedure of cleave-and-analysis. For more complex reactions, the technique ofHigh Resolution Magic Angle Spinning NMR (HRMAS) [2± 4] is emerging as a new tool that ® nally couldsolve the analytical diæ culties of SPOC. With exactly thesame pulse sequences as in high resolution liquid NMR,time requirements and resulting resolution can equally becompared. We will discuss in this lecture our approach ofHR MAS NMR, including (1) the diå usion ® lter wedeveloped to allow work in protonated organic solvents

[5], (2) diå erent schemes to monitor reaction completion[6], (3) questions of impurity detection [7] and (4) theextension to macroscopic supports that make samplehandling and hence automatic analysis more easy [8].A number of examples involving less studied chemistrieson the solid state will be shown to demonstrate thepractical feasibility of the method.

1. Baldino, C. M., Journal of Combined Chemistry, 2 (2000), 89.2. Fitch et al., Journal of Organic Chemistry, 59 (1994), 7955.3. Anderson et al., Tetrahedron Letters, 36 (1995), 5311.4. Pop et al., Tetrahedron, 52 (1996), 12209.5. Warrass et al., Journal of the American Chemical Society, 121 (1999),

3787.6. Warrass et al., Journal of Organic Chemistry 2000, 65, 2946.7. Rousselot-Pailley et al., Tetrahedron, 56 (2000), 5163.8. Rousselot-Pailley et al., Journal of Combined Chemistry, 3 (2001),

559.

Compliance and the automated pharmaceuticallaboratory

Kyle McDu¤e, CSols, Inc., 7600 Southland Blvd, Orlando, FL32809-6975, USA

21 CFR Part 11 Electronic Records, Electronic signa-tures has provided an opportunity for dramaticallyincreasing the level of information management automa-tion used in Pharmaceutical R&D and Quality Assur-ance laboratories. Although LIMS have been in use formany years, most laboratories still maintain laboriouspaper transcription and review processes for analyticalresults. Results from instruments are printed on paper,pasted in notebooks and then entered into the LIMS.Once entered, they are printed again by supervision forreview and then signed (and approved in the LIMS aswell). Next generation instrument integration tools pro-vide the level of automated work¯ ow that are needed bylaboratory and Quality Assurance staå to eliminate theneed for paper calculation and transcription steps. Theseinclude typical calculations needed for content unifor-mity, dissolution and potency. In addition, they providethe level of robustness and regulatory compliance neededto defend the quality of the laboratory data. Thispresentation will focus on the key elements of 21 CFRPart 11 which apply to laboratories operating in GMPenvironments. Successful current and future approachesto addressing the compliance, quality and work¯ owautomation challenges will be discussed.

Speciation of arsenic, selenium and antimonyusing HPLC: atomic � uorescence spectrometry

Derek W. Bryce, Warren T. Corns and Peter B. Stockwell, PSAnalytical Ltd, Arthur House Cray¢elds Industrial Estate, MainRoad, Orpington BR5 3HP, UK

The speciation of arsenic, selenium and, to a lesser extentantimony, has gained in importance over the last decade.Diå erences in toxicity, bioavailability and integration ofthe diå erent species in the biochemical cycle make theneed for accurate speciation data imperative for furtherunderstanding. Arsenic is normally found as toxic inor-ganic forms in the environment, which may undergobiomethylation by microorganisms to relatively non-toxic


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organic forms such as DMA, MMA and arsenobetaine.Selenium’ s dual nature as both an essential and toxictrace element has been widely reported, but morerecently it has been linked with cancer prevention whenfound as selenoamino acids. Antimony emissions fromanthropogenic sources such as traæ c and the productionof storage batteries has lead to an increase in antimonylevels in the environment where SbIII is less toxic thanSbV, although organic antimony species may also befound. Speciation of these elements may be carried outusing a variety of approaches, generally coupling aneæ cient separation technique with an element speci® cdetector. Various HPLC approaches have been used,either using strong anion exchange or ion-pairing HPLCto separate the species of interest. Atomic ¯ uorescence isideal oå ering unrivalled sensitivity, linearity and freedomfrom interferences, making it ideal for speciation studies.Results will be reported for the speciation of the targetanalytes including information on separations and postcolumn hydride generation. On-line oxidation and re-duction of non-hydride forming species such as arseno-betaine and selenate will be discussed and applicationsfor various samples will be shown.

Zeeman atomic absorption spectroscopy withhigh-frequency modulation with a thin-walled me-tallic hollow cathode as a base of a new mobilemonitoring system

Victoria S. Vergizova, Alexander A. Ganeev, Stanislav A.Suprunovich, Alevtina A. Kovaleva, Ilya V. Shuvaev, St-Peters-burg State University, Chemistry, Universitetskij, St Petergo¡ St-Pete 198904, Russia

Nowadays, one of the most actual issues is the develop-ment of mobile system and tools allowing one to analyseheavy metals in a monitoring regimen. Such a tool shouldbe of high analytical characteristics like traditionalanalytical techniques (high sensitivity, low detectionlimits) and provides mobility and besides monitoringanalysis (low applied power, small size of technique,moreover absence or minimum of chemical samplespretreatment are desirable). One of the way to realizeall of these features is to develop technique based on thin-walled metallic hollow cathode (TMHC) as atomizerwith high-frequency modulation Zeeman atomic absorp-tion spectroscopy (providing high sensitivity and analysisof complex samples without or with minimal samplepretreatment). In this technique, dry residue of solutionis analysed. Owing to thermal-ionic mechanism and longtrap time of analysed atoms in ` analytical signal formingzone’ observed in TMHC low detection limits and highsensitivity compared with ones in graphite furnace takeplace. Moreover, due to self-clearing eå ect under dis-charge process TMHC has extremely low memory eå ect.Applied power is about two orders of magnitude lowerthan required to analyser with graphite furnace, dis-charge gas (Ar) consumption is also extremely low. Thereis possibility to analyse 2500 samples without TMHCchanging in contrary to graphite furnace providing aboutorder lower resource. All these advantages have allowedus to design new mobile technique allowing monitoringanalysis of metals even in complex environmental and

biological samples. Analytical parameters of new tech-nique such as detection limits and matrix eå ects arepresented. Detection limits for Cd, Mn and Cu are 0.3,3 and 6 pg accordingly.

Electronic nose as a quality-control tool in thepharmaceutical industry

Jean-Christo Mifsud and Sophie Puech, ALPHA M.O.S, 20Avenue Didier Daurat, Toulouse F-31400, France

Flavours qualities and intensities attract an increasinglevel of attention from the pharmaceutical companies,particularly the one involved in the development ofpaediatric formulations. This industry invests heavily ininstruments that can help it to ensure the quality and theconsistency of their product. Diå erent qualities andintensities of various ¯ avours used in pharmaceuticalformulations were successfully evaluated using aFox4000 Electronic Nose (Fox4000 EN). The electronicnose has generated great interest in the analyticallaboratories of the world’ s leading food, ¯ avours, andfragrance companies as a fast, simple, and reliablemethod of aroma/VOC analysis. Over the past 5 years,the sensors arrays system have demonstrated their abilityto produce information (instead of a stream of data) andthe ability to transfer expert knowledge from the R&Dand trained sensory panels into a production environ-ment for quality assurance and control. Good discrimi-nation and excellent reproducibility were obtained. Thesuccessful identi® cation of acceptable quality and inten-sity of ¯ avours in ` blind tests’ have been obtained onunstressed and stressed samples for shelf life evaluation.Interbatch training sets of ¯ avours were used to ensurethat a representative batch variation was introduced intothe model using custom identi® cation method and also toensure that the discrimination level of the acceptable,marginal and unacceptable ¯ avours were suæ cient whendiå erent batches were considered. Both Principal Com-ponent Analysis (PCA) and Discrimination FunctionAnalysis (DFA) were used for data processing andidenti® cation. Good correlation between GC data, sen-sory panel and the Fox4000 EN were found. Finally,the principle of overall ¯ avours intensity measurementwas demonstrated using Partial Least Square (PLS) byshowing data for ¯ avours from various batches of recep-tion and various intensities.

A multichannel biochip with bio� uidic module forreal-time bioassays

David L. Stokes, Guy D. Gri¤n, Leonard R. Allain, Dimitra N.Stratis and Tuan Vo-Dinh, Oak Ridge National Laboratory,Advanced Monitoring Development, 1 Bethel Valley Road, OakRidge, TN 37830-8050, USA

This work describes the development of a bio¯ uidicssystem that is coupled to a multichannel biochip sensorfor real-time monitoring of bioassays. The bio¯ uidicsmodule features a single reaction chamber in whichmultiple bioassays can be performed simultaneously. Inthis technique, a sampling platform (e.g. membrane) isprepared oå -line with a modi® ed ink jet printing tech-nology which allows rapid array printing of multiple,


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discreet capture probe zones or microdots on a singlesampling platform. The sampling platform is then placedin the reaction chamber of the bio¯ uidics module, andthe pattern of capture probe microdots is aligned with anarray of independently addressable biochip photosensors .The bio uidic system delivers samples and reagents forthe ensuing bioassays. The system can accommodateboth DNA and antibody-based assays, both of whichhave been performed on a single sampling platform. Thereaction chamber also features a nichrome heating ele-ment and thermocouple probe for performing on-chipreactions at optimized temperatures. The bio¯ uidics-based multichannel biochip detector has been appliedto the detection of a variety of pathogens and analogues,including E. coli and B. globigii.

Development of a semi-automated comprehensiveextraction and multiple fractionation (S-ACEMF)METHOD—Part I: Evaluation of di erent solidphase sorbets and optimization of overall recov-ery

Courtney D. Sandau, Andreas Sjodin, Mark D. Davis, Alyson L.Waterman, William Roman and Donald G. Patterson, Centersfor Disease Control and Prevention, Toxicology Branch, 4770Buford Hwy 1 Mai, Atlanta, GA 30341-3717, USA

A semi-automated extraction method has been developedto increase our current sample throughput. The methodemploys solid phase extraction with automation onthe RapidTrace1(Zymark Corporation). The currentmethodology has been evaluated for polychlorinatedbiphenyls (PCBs), hydroxylated PCB metabolites,methylsulphonyl PCB metabolites, persistent pesticides,polychlorinated naphthalenes and polybrominateddiphenyl ethers. Sorbent evaluation: Nine diå erentsorbents from several manufacturers were evaluated,which included C181, ENV1, NEXUS1(all Varian,Inc.), OASIS1 (Waters Corporation) and Chroma-bond1 (Macherey-Nagel) . Cartridges of all sorbentswere packed in house to correspond to bed heights of4, 12, 22 and 30 mm, respectively, in 3-ml cartridges.The weight of the sorbents used to obtain a certainbed height varied due to diå erent densities of thesorbents. Duplicate samples were extracted for eachbed height, using the RapidTrace1, employing the sameextraction procedure. The procedure included (1)conditioning, (2) application of sample, (3) drying ofcartridge and (4) elution of compounds of interest.All serum samples (2 ml) were pretreated using thesame methodology employing formic acid (2 ml),sonication, and subsequent dilution with water (2 ml).Sorbent heights were plotted against absoluterecoveries for each sorbent and analyte. Recoveriesfor highly lipophilic compounds such as decachloro-biphenyl (CB-209) were lower than recoveries forless chlorinated and less lipophilic compounds, suchas 2,2,3,30,4,40-hexachlorobiphenyl (CB-153). However,recoveries increased for CB-209, to approximately80% using a bed height of 30 mm for most sorbents,indicating how important these experiments are fordetermining the amount of sorbent needed for eæ cientextraction.

Development of a semi-automated comprehensiveextraction and multiple fractionation (S-ACEMF)method—Part II—Isolation and puri� cation ofPBDES, persistent pesticides, PCBs and PCBmetabolites from serum

Courtney D. Sandau, Andreas Sjodin, Mark D. Davis, Alyson L.Waterman, William Roman and Donald G. Patterson, Centersfor Disease Control and Prevention, Toxicology Branch, 4770Buford Hwy 1 Mai, Atlanta, GA 30341-3717, USA

The comprehensive extraction method described bySjodin et al. was coupled with a multiple fractionationmethod to allow the separation and quantitation ofpolybrominated diphenyl ethers (PBDEs), polychlori-nated biphenyls (PCBs), persistent pesticides, haloge-nated phenolic compounds (including hydroxylatedPCBs), and methylsulphonyl-PCBs (MSF-PCBs). Eachof the fractions were isolated using speci® c interactionswith diå erent commercially available sorbents and weresemi-automated using the Zymark RapidTrace1system(Zymark Corporation). The serum extract from thepolymeric extraction sorbent was reduced in volumeand separated into two fractions on activated (100%)silica gelÐ non-polar compounds (F1) and polar com-pounds which includes phenolic compounds, MSF-PCBsand polar pesticides (F2). Phenolic compounds in F2were then derivatized to their methyl ethers and cleanedup further on silica/sulphuric acid columns to removeresidual biogenics before mass spectral analysis. The non-polar fraction (F1) can be fractionated further on acarbon column to separate PCBs/persistent pesticides(F1a) from planar compounds, such as polychlorinatednaphthalenes, dioxins and furans (F1b). Both fractionswere puri® ed when necessary before gas chromatogra-phy-mass spectral analysis. The ® nal semi-automatedmethod will be described along with the application ofthe method to human serum and plasma. The ZymarkRapidTrace1allowed semi-automation of a complexmethod that signi® cantly increased throughput ofsamples in our laboratory and also increased the numberof analytes monitored due to the fractionation techniquesdescribed above. The potential of this method to replacethe current vacuum manifold methods and to expandfurther to other classes of compounds, such as dioxins,furans and polychlorinated naphthalenes will also bediscussed.

Development of an integrated electrochemica l im-muno-analysis on microchip platforms

Madhu Prakas Chatrathi, Joseph Wang, Alfredo Ibanez andAlberto Escarpa, New Mexico State University, Department ofChemistry and Biosciences, MSC 3C North Horse Shoe Drive,Las Cruces, NM 88003, USA

Micro¯ uidic devices have achieved rapid development inthe recent years and continue to advance toward fullyautomated tool. The power and scope of micro-analyticalsystems can be greatly enhanced by performing highlyselective biochemical reactions, in particular, on-chipimmunoassays using highly speci® c antigen± antibodyinteractions. Previously reported microchip based immu-noassays have been performed using either thermal lensmicroscopic (TLM) or laser induced ¯ uorescence (LIF)


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detection methods, however, there are no reportsof analogous on-chip electrochemical immunoassays .One of the bene® ts of using electrochemical detectionwith microchips is the ability to maintain good detectionlimits with other bene® ts such as tuneable selectivity,high sensitivity and compatibility for further miniatur-ization. The present investigation focuses on a completeintegration of individual steps of electrochemical immu-noassay on chip. This includes precolumn reaction ofalkaline-phosphatas e labelled antibody with the antigen,electrophoretic separation of free antibody from anti-body± antigen complex, followed by amperometric detec-tion of 4-aminophenol product resulting from thereaction of 4-aminophenyl phosphate substrate with theenzyme tracer (alkaline phosphatase) in the post-column.In this paper, analytical protocols are described, con-ditions are optimized and data will be presented quanti-fying the performance. A remarkably low detection limitof 2.5 £ 10¡16 g ml¡1 is obtained for the mouse IgGmodel analyte. This approach has a universal applic-ability as the target analyte of the assay can be changedsimply by changing the antibody and is limited only bythe availability of antibody for the analyte of interest.

Chemometric analysis of two-dimensiona l gaschromatographic data

Kevin J. Johnson, Bryan J. Prazen and Robert E. Synovec,University of Washington/Center for Process Analytical Chem-istry, Department of Chemistry, Seattle, WA 98195, USA

Two-dimensional gas chromatography (GC £ GC) isapplied to both a pattern recognition problem involvingclassi® cation of jet fuel mixtures and to a trilinear partialleast squares (tri-PLS) quanti ® cation of the compositionof industrial naphtha samples. The use of two chromato-graphic columns with complementary retentive proper-ties in GC £ GC separations results in a signi® cantincrease in both peak capacity per unit time andselectivity over separations obtained with single-columnGC methods. High-speed GC £ GC analysis is accom-plished through the use of short GC columns and highgas velocities as well as through the practice of followingpartial chromatographi c peak resolution with multivari-ate chemometric analysis. This work is aimed at provid-ing a GC system for rapid quantitative and qualitativeanalysis of complex samples such as process streams. Injet fuel classi ® cation studies, jet fuel samples are subjectedto a high-speed 5-min GC £ GC separation. An analysisof variance (ANOVA) based feature selection method isthen used to identify and select chromatographic featuresrelevant to a given classi ® cation between mixtures ofdiå erent jet fuel types. In one experiment, 1% volumetriccompositional changes in binary mixtures of JP-5 and JP-7 jet fuels are readily observed in scores plots fromprincipal component analysis (PCA) of the selectedfeatures. In a second experiment, features adept atclassi® cation of jet fuel type but not sensitive to geo-graphic origin of the sample are located in a data setconsisting of GC £ GC chromatograms of samples ofthree diå erent jet fuel types. In the naphtha analysisstudy, trilinear partial least squares models are used topredict the aromatic and naphthene content of in-dustrial naphtha samples. The tri-PLS models are con-

structed using chromatograms from six minute GC £ GCseparations of neat naphtha samples and correspondingreference values that were determined by a standardsingle-column GC method. The GC £ GC/tri-PLS analy-sis method is shown to provide acceptable quantitativeprecision while using a separation time that is more than15 times faster than the single-column standard method.

Sample management for DNA testing laboratories

Nick Townsend, Martin Waugh and Peter B. Mans¢eld,Autoscribe Ltd, 81 Technology Park Drive, East Falmouth,MA 02536, USA

DNA technology has revolutionized applications such aspaternity testing, forensics, clinical diagnosis and thedetection of genetically modi® ed organisms (GMOs).This success in turn has created a very large increase inthe number of samples being processed by DNA testinglaboratories and the associated logistical issues this canbring. In the last two decades LIMS have becomeestablished supporting the work of analytical and re-search laboratories but is it only recently that DNAtesting laboratories been able to realize the bene® ts thatLIMS can deliver. In this presentation we will describe aLIMS con® gured to meet the needs of DNA contracttesting laboratories. We will describe how, via Webtechnology, clients submit samples for testing, specifythe DNA testing pro® le and con® rm order details. Oncesamples are received, bar code technology is used to tracksample location and process steps including extraction,puri® cation, ampli® cation and identi® cation. A verygraphical and intuitive interface is provided for techni-cians allowing creation and editing of plate and electro-phoresis gel plans using drag-and-drop techniques. Thesystem supports the use of complex ` plating rules’ provid-ing a large degree of automation as to how primers,samples and reference standards should be pipetted ontothe plates. A comprehensive audit trail is providedrecording details of how samples are analysed includingthe run conditions associated with processing the platesand the transfer of samples from plates to gels. Onceanalysis is complete the LIMS automates the productionof reports for the client and will interface with orderprocessing systems to deliver an invoice for the work.

Automated sample spotting on MALDI plates

Alan Hamstra, Tim Hegeman and Luke Roenneburg, Gilson,Inc., 3000 W. Beltline Hwy, Middleton, WI 53562-1617,USA

Sample preparation is a rate-limiting step in the analysisof samples with MALDI-TOF instrumentation. Platepreparation requires adding both sample and matrix,allowing them to form a homogenous mix and thendrying. Knowing the limitations of hardware, sampleconcentration, sample volume and sample density helpspredict sample thoughtput. Hardware is available withpositional accuracy to < 100 mm, however positionalaccuracy is only one parameter aå ecting the sample areaoccupied on a plate. MALDI plates come in a varietyof sizes and con® gurations, placing requirements onsample application hardware. The volume of available


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sample imposes additional limits on hardware. The cal-culated theoretical diameter occupied by a 100 nlsphere is 726 mm. When placed on the plate surface, thedrop compresses vertically and occupies a diameter> 726 mm. Plate surface chemistries, either hydrophobicor hydrophilic, also aå ect plate sample density. Matrixand sample surface tension and viscosity aå ect plate areaoccupied by sample. To pass through the small probe, theori® ce matrix and sample must be free of any particulateor a probe will plug and dispense inaccurately. Requiredsample accuracy and carryover speci® cations place timelimitations on plate preparation. Loading a 384-wellplate with matrix requires only minutes, while rinsingprobes between samples to prevent carry over requiresfrom 20 to 60 min. A rinse cycle may take signi® cantlylonger than sample application and account for 50± 90%of the time required to load a plate. Technology isavailable to dispense nanolitre volumes of sample, how-ever there are limits on the sample volume, which can beaspirated. If the minimal aspirated sample volume islimited to microlitres, each dispense of nanolitres maywaste microlitres. Hardware and software must recognizethis limitation and take steps to conserve sample. I willpresent data on minimizing the area occupied by sample,optimizing sample volume and mass, and minimizingplate preparation time.

Millisecond monitoring of fast transient signalsby Hadamard transform time-of- � ight mass spec-trometry

Joel R. Kimmel, Facundo M. Fernandez, Jose M. Vadillo andRichard N. Zare Chemistry Department Mudd Bldg, 333Campus Drive, Stanford, CA 94305-4401, USA

Hadamard transform time-of-¯ ight mass spectrometry(HT-TOFMS) involves the modulation of a continuousion beam using a pseudo-random sequence of pulses [1].This sequence of pulses is applied using a chargedparticle modulator consisting of an interleaved combof wires. Ions ¯ ying from the electrospray ion sourceto the ¯ ight chamber are ` gated’ at this modulatorfollowing the Hadamard-type binary sequence. Theapplied sequence has an approximately equal numberof zeros and ones, thus the duty cycle of thespectrometer is close to 50%. TOFMS has become thepreferred detector for separation techniques used inbioanalytical chemistry such as CE and LC. As researchcontinues, the obtained separation eæ ciency of thesemethods increases, making the eluting peaks sharper.To describe correctly the shape of these narrow peaks,detectors must acquire data at a rapid speed. Theonly means of increasing the spectral acquisition speedof a TOFMS without sacri® cing mass range is touse encoding methods that eliminate the lag timebetween successive ion packets. In this presentation,we describe the ability of HT-TOFMS to scan transientsignals in the millisecond time-scale. A brief theoreticalbackground on how TOFMS can be multiplexedis given. Alternative Hadamard transform algorithmsthat improve the performance of slow multichannelscalers are discussed in terms of their eå ect onthe spectral storage rate. The eå ect of acquisitionsystems’ end-of-pass dead time is examined. The

ability of HT-TOFMS to scan transient peaksof decreasing width is investigated for CE and pulsedelectrospray ionization peaks. For femtomoleinjections of biomolecules, full spectra in a mass rangeof 1500 Da are obtained in 3.6 ms. Owing to theextremely large amounts of data collected, only state-of-the-art acquisition systems can be used for this task.Further increase in the spectral storage rate can only beachieved by further increasing the duty cycle of theinstrument. The possibility of using two MCP detectorsto produce a 100% duty cycle with no ion storage isdiscussed.

1. Zare et al. Journal of the American Society of Mass Spectrometry, 12(2001), 1302.

Quasi-continuous monitoring of process andwaste streams by on-line inductively coupled plas-ma-optical emission spectroscopy

Dirk Ardelt, Heinz Falk, Hendrik M. Smol and Hans-JoergWaarlo, Spectro Analytical Instruments GmbH & Co. KG,Product Group ICP (T-3), Boschstrasse 10, Kleve D-47533,Germany

Over the past decade, increasingly more sophisticatedprocess analysers derived from ` mature’ laboratorymethods of instrumental analysis have found widespreaduse in the chemical process industry to cope withincreasing demands for more eæ cient energy andmaterial usage and better process control. Althoughmany such applications still rely on rather simple sensors,an increasing amount of formerly laboratory based onlymethods, like infrared or Raman spectroscopy, X-ray¯ uorescence or magnetic resonance methods, arenowadays routinely applied as on-line systems. As diverseas these methods are, their ® elds of application are inthe on-line process environment, which typically isdivided among the distinction between the aggregatestates of the medium to be analysed. Not surprisingly,one major ® eld is the analysis of liquid process and wastestreams. Considering laboratory use, inductively coupledoptical emission spectroscopy (ICP-OES) probably is themost universal atomic spectroscopy method existing.Combining high ¯ exibility, good sensitivity, widedynamic range and fast measurement cycles, this methodshould ideally be suited for migrating into the processanalytical ® eld, especially regarding its relative ` matur-ity’ . Despite of these ® ndings, ICP-OES up to now hasnot found the expected use as on-line analyser, evenwhere it outperforms other methods under laboratoryconditions. Using recently installed systems forwastewater and process stream monitoring as examples,we will discuss probable reasons for the rather slow pastdevelopment of on-line ICP-OES and examine how farthese reasons have been or can be overcome. Relevantanalytical and technical ® gures of merit for on-lineapplications shall be deduced. Furthermore, modernmultichannel detection ICP-OES allows for usingsophisticated algorithms (e.g. chemometrics) for dataevaluation, in this case resulting in higher degrees ofautomation and system availability. Accounting for suchadvances, we will try de® ning a ` state of the art’ for ICP-


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OES as process analytical method and highlight futuredevelopment areas.

A new stand-alone autosampler for high-through-put HPLC and HPLC/MS

Curtis R. Campbell, Susan M. Steinike and Kara M. Merkle,Shimadzu Scienti¢c Instruments, LCMS Applications, 7102Riverwood Drive, Columbia, MD 21046-1245, USA

Fast LC and fast LCMS methods have been developed tomeet the demands of combinatorial chemistry and drugdiscovery. Now more than ever is there a correspondingrequirement for ultra-fast sample introduction in LCand LCMS instruments along with the most stringentrequirements for zero carryover and optimum samplerepeatability. In this communication we describe the per-formance of a newly designed stand-alone autosamplerfor HPLC and HPLC/MS applications featuring a 15-sinjection cycle for 10-ml injections, excellent samplerepeatability and the lowest carryover currently attain-able. The unit can function independently, allowinginterface with a variety of instrument models in a varietyof venues including combinatorial chemistry, drug dis-covery and high-throughpu t screening.

Ultra-fast liquid chromatography: the importanceof system temperature

Jon D. Thompson and Peter W. Carr, University of Minnesota,Department of Chemistry, 207 Pleasant Street SE, Minneapolis,MN 55455-0431, USA

It is very important that the speed of HPLC be increased.Improving throughput for stability analysis, qualitycontrol assays and dissolution testing are all keymotivations for improvement in separation speed inthe pharmaceutical industry. Unlike CE, HPLC doesnot lend itself well to multiparallel analysis becausethe cost is prohibitive. Recent work in this laboratoryhas focused on the theory of high temperature onanalysis time in LC. Disregarding selectivity con-siderations, we have shown that the 5± 10-fold decreasein eluent viscosity that comes from working at veryhigh temperature (180± 200 8C), and the concomitantincrease in analyte diå usitivity, combine dramaticallyto decrease the time needed to generate a theoreticalplate. The lower viscosity at elevated temperaturedecreases the pressure drop across the column andallows higher linear velocities to be reached as thepump’ s pressure limit is approached. Simultaneously,faster analyte diå usion at higher temperature improveseæ ciency at high linear velocity conditions comparedwith the eæ ciency at lower temperatures at the samevelocity. In this presentation, we will show how thetheory of speed guides column selection and systemdesign. We will show that at high temperatures, 10gradient runs can be done in 40 min on columns ofconventional geometry. We will also show that dramaticimprovements in stabilityÐ indicating assay through-putÐ can be achieved at elevated temperature usingconventional equipment.

Development of a high-throughput analysis infra-structure for combinatorial materials science ap-plications

Radislav A. Potyrailo, General Electric, Corporate Research andDevelopment, PO Box 8, Schenectady, NY 12301-0008, USA

High-throughput methods for materials science combinea parallel or combinatorial materials synthesis andprocessing with an automated screening and datamanagement tools. They can rapidly optimize molecularproperties and process conditions that are diæ cult topredict using existing knowledge. From an analyticalchemistry perspective, this leads to major challenges indeveloping rapid analysis techniques that can deal withlarge numbers of small samples required to constructcombinatorial libraries. To meet these analyticalchallenges in the discovery of new materials at GeneralElectric, a high-throughpu t analysis infrastructurewas established at GE’ s Corporate Research Center.This presentation will describe the strategy taken inthe development of this infrastructure. For high-throughput analysis of materials properties from a widevariety of projects, analytical instrumentation consistsof interchangeable modular subsystems coupled withthe new data-processing capabilities. Examplesfrom several projects will illustrate the broad applic-ability of the developed high-throughput analysisinstrumentation.

High-throughput analytics employing automatedsample preparation and the integrated use ofHPLC-DAD, HPLC-DAD-MS, GC-MS and � ow in-jection NMR

Winfried Etzel1, Stefan Beeck1 and Wolfgang Gau2, 1BayerAG, Agrochemical Division, Chemical Research GEB 6530,Leverkusen D-51368, Germany, 2Bayer AG, Landwirtschaftszen-trum Monheim, Alfred-Nobel-St r 50, 6530, Monheim D-40789,Germany

The establishment of automated and parallelizedsynthesis and micro-scale reactions in the chemicalsynthesis laboratories of the Agrochemicals Division atBayer created an increasing demand for analyticalinformation (purity, structure, physicochemicalproperties). With the availability of ¯ ow injectionNMR systems (BEST-NMR) the use of the samesample format on diå erent analytical instruments waspossible. This fact encouraged us to realize an integrationof diå erent analytical techniques (NMR, HPLC-DAD,HPLC-DAD-MS, GC-MS) in one service unit.The optimization of the ¯ ow NMR system for theuse of diå erent organic solvents will be demonstratedand the quality of the spectra and the throughput ofsamples discussed. In addition, the results andadvantages of an integration of diå erent analyticaltechniques will be presented. Working with barcodesand microtitreplate (MTP) technology were introduced.Electronic laboratory journal, sample preparationrobotic system and analytical instruments wereconnected together via a laboratory information andmanagement system (LIMS). This resulted in asimpli® ed work¯ ow in the chemical research laboratoriesand in the analytical department. The throughput


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of samples and measurements was doubled withoutraising the number of technicians.

An automated system for combined samplepreparation and analysis for high-throughput-screening

Kerstin Thurow, Agnes Schubert and Christian Wendler, Uni-versity Rostock, Institut Automatisierungstechnik, R-Wagner-Strasse 31, Rostock-Wamemunde D-18119, Germany

The use of combinatorial methods in chemistry andlife science has been developing rapidly within the lastyears. One of the ` bottlenecks ’ in synthesis control inpharmacy and life sciences is still the characterizationand speciation of biotechnological reaction productsusing chromatographic methods in combination withspeci® c detection systems. Often before analysis, a com-plex sample preparation is required which might includecleaning steps, dilution or even derivatization pro-cedures. Existing systems are not ¯ exible automatedsolutions and do not have ¯ exible material and informa-tion interfaces. Thus, the objective of our investigationwas the development of an automated system for com-bined sample preparation and analysis being used fororganic synthesis. The system used is a commercialliquid-handling station (CTC CombiPal) which has beenequipped with diå erent trays for heating, ® ltration,mixing etc. and has been adapted to diå erent analyticaldevices such as gas or liquid chromatographic systems ormass spectrometers. Technically, the system is controlledby a digital computer operating both chromatographand sampler as diå erent tasks in the WINDOWS-NT 4.0operating system. The sample preparation task allowsthe application of user speci® c methods. Sampler andchromatograph communicate for transmitting remoteSTART/STOP-information via hardware handshakelines. Because of the ¯ exible system strategy, the systemcan be set up easily for diå erent applications. The systemdeveloped allows the fully automatic analysis of samplesto run in parallel with the sample preparation. It can beused as a stand-alone system or can be integrated incomplex robotic systems. Motivation for such a combina-tion is to reduce testing costs further by increasingthroughput and reducing manual intervention. Otheradvantages include, for example, reducing the delaybetween the analysis and the sample preparation. Thepresentation will outline the technical details of thesystem developed and will show the use of the systemfor high-throughput screening applications in the ® eld ofcombinatorial catalysis, chiral determinations and syn-thesis control.

Modi� cation of OSHA GC methods for continuousarea monitoring

John N. Driscoll, Process Analysers, LLC, 25 Walpole ParkSouth Drive, Walpole, MA 02081, USA

OSHA has developed hundreds of GC methods foranalysing organic compounds in the workplace as partof the standards process. These methods were developedto provide analytical methods for samples collected onadsorbent tubes in the workplace for a wide variety of

industries. Typical sample volumes collected range from250 ml (at the ceiling value) to 12 litres (below the TLV)of sample collected on the adsorbent tubes. Once thesample (250 ml) is diluted, it is equivalent to injecting a1 cm3 sample at approximately 5£ the TLV. For areamonitoring, we would like to have a detection limit thatis 1/20th of the TLV or PEL. If these detectors were usedin an Automatic GC for Area monitoring, many wouldhave diæ culty detecting half the TLV for those com-pounds with low TLVs. In other words, many of thesemethods would be ` sensitivity challenged’ because ofthe use of the ¯ ame ionization detector (FID). TheseOSHA GC methods can still be used but it is clear thata more sensitive method (or a concentrator for theFID) is needed to obtain the required sensitivity. Thephoto-ionization detector (PID) is > 50 times moresensitive than an FID for aromatic compounds. This willallow a smaller sample (0.1± 3 cm3) to be used and stillhave adequate sensitivity for the method. In addition, theselectivity of PID can be varied by choosing a 9.5, 10.2 or11.7 eV lamp. The sensitivity of the FID can be increasedvia an on-board automatic concentrator that can im-prove the detector sensitivity by 10± 100-fold. The com-pounds to be concentrated can be optimized by switchingthe trap materials. Another diæ culty that occurs at lowppm or ppb levels with polar species is adsorption orreactivity on/with surfaces of the sampling system andlines. This results in serious problems with reproducibilityand accuracy. Examples include amines, phenols, organ-ic and inorganic acids, pesticides, etc. A permeation tubeor other diå usion device can be used to condition theentire system and prevent adsorption of polar or reactivecompounds. We will evaluate a number of methods foramines, sulphur compounds, chlorinated hydrocarbonsand diphenyl oxide. We will compare these results of theFID/NPD detectors with the photo-ionization detector(PID) mounted on an automatic GC for area monitor-ing. This should provide an alternate method that hasmore sensitivity and/or selectivity and does not requiresupport gases.

Fast gas chromatography with conventional in-struments using direct resistively heated capillarycolumns

Paolo Magni, Giacinto Zilioli and Riccardo Facchetti, Thermo-Quest Italia SpA, Research and Development, Strada Rivoltana,Rodano, Milan I-20090, Italy

In gas chromatography , there is an increasing demandfor signi® cant reductions in analysis time. Howeverperforming high-speed separations while keeping enoughseparation eæ ciency is a diæ cult task. The combined useof fast temperature programming and short narrow-borecolumns may provide an optimal solution, particularlyfor samples containing simple mixtures with a wideboiling point range. For some applications a suæ cientseparation can be achieved even in < 1 min providedthat, in the same time, the column is raised to anappropriate ® nal temperature. The system presented inthis paper, consisting in a directly heated capillarycolumn module mounted in a conventional GC instru-ment, permits one to achieve very fast temperatureprogramming rates (as high as 20 8C/s), which cannot


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be obtained with the use of conventional air circulatingGC ovens. The tremendous heating rate power of thedevice permits one to reduce the tight bandwidth sampleinjection requirements otherwise dictated by the smallinternal diameter of the column. The module, containingthe capillary column, the heating element and thetemperature sensor, is housed in the GC oven andconnected to standard GC split-splitless injector anddetection systems, just as any conventional capillarycolumn. Results obtained with both split and splitlessinjection techniques are examined using Flame Ioniza-tion Detector and Time of Flight Mass Spectrometer.The attached chromatogram shows an example of famesanalysis, ranging from n-C7 to n-C22 frames, performedin < 1 min. The separation was obtained using a 5 m,0.1 mm i.d., 0.1 mm ® lm thickness RTX-5 column with atemperature-programmin g rate of 5 8C/s (from 70 to300 8C). Others applications of the fast GC accessory inthe chemical, petrochemical, environmental, and foodand ¯ avours ® elds are presented.

Screening for environmental contamination usingliquid chromatography with mass spectrometrydetection

Jim Krol and Kate Yu, Waters Corporation, 34 Maple Street,Milford, MA 01757-3696, USA

When assessing unknown contamination of anenvironmental sample, where does the chemist begin?Which validated method is appropriate, a speci® canalyte method or several diå erent analyte methods?What if the validated method reports no contamination;productivity decreases, analysis cost increase and thecontamination analyte remains unknown. Liquidchromatograph y (LC) has the capability to retainand separate numerous analytes based upon theirchemical properties, but conventional PDA-UVdetection has limitations in seeing all the analytes. Foranalyte identi® cation, retention time alone is notsuæ cient. Chromatographic analyte coelutions willexist in complex samples and compromise UV spectramaking identi® cation and quantitation marginal. If theanalyte is UV inactive, analyte identi® cation andquantitation are impossible. Mass spectrometry (MS)is a more universal, yet speci® c detection techniquethat detects a signi® cantly greater number ofenvironmental analytes, such as carbamate and ureabased pesticides and herbicides using positive electro-spray. Combined with gradient reverse phase chromato-graphy, an environmental sample can be screened fornumerous analytes. The simultaneous use of retentiontime, m/z mass spectra and PDA-UV are used asqualitative variables for analyte identi® cation. However,the low ppb semiquantitative results need to be con-® rmed by speci® c validated methods. This presentationwill present a novel carbamate and urea analyte screen-ing method approach. The critical link is the reliability ofthe MS processing method, chromatographic reproduci-bility, and the percentage of false-positives/false-nega-tives. Wastewater and ground water will be theevaluated matrices.

Miniaturized � ow-through surface plasmon res-onance detector for the study of protein–proteininteraction dynamics

Rebecca J. Whelan1, Thorsten Wohland1, Richard N. Zare1,Lars Neumann2, Jacqueline Steenhuis2 and Brian Kobilka2,1Department of Chemistry, Mudd Chemistry Building, Stanford,CA 94043, USA, 2Departments of Medicine and Cardiology,Beckman Center, Stanford, CA 94305, USA

Surface plasmon resonance (SPR), which senses changesin the refractive index of a dielectric medium adjacent toa thin gold ® lm, is a powerful tool for label-free studies ofbiological molecules and their interactions in real time.We employ SPR to study the interaction of theG-protein-coupled beta-2 adrenergic receptor with othermolecules, including receptor agonists and antagonists ,kinases, and G. protein. The most relevant informationcomes from receptors that are immobilized on the surfacein a uniform way. To this end, we have developed a novelmutation of the receptor, with an additional cysteine atthe extracellular terminus. Biotinylation of this mutantreceptor and use of conventional biotin/avidin binding® xes the receptor to the gold ® lm of the SPR sensor withexcellent uniformity. An alternate immobilizationprotocol uses an antibody (M1) directed against thereceptor’ s amino terminal FLAG tag. The receptorsretain their function after immobilization, as con® rmedby ¯ uorescence microscopy studies. After introduction ofa reaction partner through a ¯ ow cell (operating with orwithout continual ¯ ow), information can be obtainedabout the extent of interaction, binding kinetics, andpharmacology. Other biological interactions we haveinvestigated include antigen/antibody, glycoprotein/lectin, and ssDNA/ssDNA. Regardless of the analytesystem, maximum sensitivity and eæ ciency are achievedby minimizing the volume of sample above the sensingsurface. To minimize the required sample, we havedeveloped a miniaturized ¯ ow cell. This small ¯ ow cellalso opens the possibility of coupling SPR with aminiaturized separation platform such as CE or micro-HPLC.

Determination of sulfhydryl residues in cysteine-rich metalloproteins using � ow-injection quartzcrystal microbalance

Alejandro Lopez Briseno, Alfred J. Baca, Fayi Song andFeimeng Zhou, California State University, Chemistry andBiochemistry, 5151 State University Drive, Los Angeles, CA90032-4226, USA

Metallothioneins (MTs) are low-molecular weight pro-teins having amino acid compositions rich in sulphurwhich strongly bind to metal ions such as Zn, Cd, Cu andHg. Extensive research has been conducted concerningthe two types of redox groups (metals and mercaptidegroups) in these molecules. Particular emphasis has beenaimed at determining the number of sulfhydryl groupsinvolved in the electron transfer reactions of metallothio-neins. However, less attention has been directed in thedetermination of sulfhydryl groups that are not directlyinvolved in the redox properties of these proteins. Wereport here our recent eå orts in determining the numberof sulfhydryl groups that directly participate in the redox


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reactions of Metallothioneins. It was found that approxi-mately four cysteines per MT molecule are involved inthe cysteine-mercury thiolate formation. This was accom-plished by combining two well-known techniques: theelectrochemical quartz crystal microbalance (EQCM)and the inductively coupled plasma atomic emissionspectrometry (ICP-AES). Another objective of this studywill be to determine the availability of non-participatingcysteine groups by signal ampli® cation through chemicalreaction to convert sulfhydryl groups into easily detect-able moieties. A conceivable method of accomplishingthis is by selective modi® cation onto cysteine moietiesacting as probes to methoxy-polyethyleneglyco l male-imide (MPEG-MAL). The quartz crystal microbalance(QCM) will be used for gravimetric analysis of thesulfhydryl groups.

Pro� ling compounds for their solubility proper-ties

Christopher A. Lipinski, P¢zer Global Research and Develop-ment, MS 8118-220, Eastern Point Road, MS 8118-2, Groton,CT 06340, USA

Oral absorption depends on adequate solubility andintestinal permeability. Poor aqueous solubility due tohigh lipophilicity is fairly easy to predict computationallyand to avoid. However, poor solubility due to crystalpacking forces is actually more common. High-through-put solubility assays can be designed in two diå erentways. In one method a DMSO stock solution of thecompound is serially diluted with aqueous medium andthe precipitation point is determined by a turbidimetricassay. An advantage is high assay capacity and theability to construct a relative solubility ranking ofcompounds. A disadvantage is the high percentage ofDMSO in the aqueous. High DMSO content also occursif 5 or 10 mm DMSO stock solutions are added to anaqueous medium. An alternative method that we usepreserves the connection to a thermodynamic solubilityassay to a greater extent. The design is based onidentifying the critical thermodynamic solubility rangefor the most commonly encountered type of compounds.Our assay mimics the early discovery method by whicha biologist orally doses an animal. Compound is addedto the aqueous as a 60 mm DMSO stock solutionthus keeping the ® nal percentage of DMSO quite low.Turbidimetric solubility values always numericallyexceed thermodynamic values, are very relevant to earlydiscovery, but should never be used in the late discoverysetting. Our two complimentary turbidimetric solubilityassays have a throughput of 10 000 compounds per year.A ¯ ow cell assay measures solubility in the range 5± 65mg/ml, the critical range for the typical poorly solubleheterocycle with average permeability and average pro-jected clinical potency. A plate reader assay measuresthe higher solubility range 50± 500 mg/ml, the critical sol-ubility range for the typical poorly permeable pep-tido-mimetic. Poor aqueous solubility has considerablerelevance to experimental permeability studies and canbe a considerable cause of experimental error even at apermeability screening dose as low as 10 mm.

Process monitoring of the moisture and � nish-on-� bre of textile products using a portable nearinfrared analyser

James E. Rodgers1, Rafael L. Barraza1, Panitan Sukpaladisai2

and Charles M. Horton3, 1Solutia, Inc., 3000 Old ChemstrandRoad, Cantonment, FL 32533-8926, USA, 2Solutia, Inc., High-way 20 West, Decatur, AL 35609, USA, 3Solutia, Inc., 1515Highway 246 S., Greenwood, SC 29646-8402, USA

The moisture content and the quantity of ® nish appliedto the surface of textile ® bres (Finish-On-Fibre , or FOF)are often critical process and quality control variables, forthey can signi® cantly impact physical properties, manu-facturing processes, quality, and productivity. A recur-ring problem for textile bobbin products is the rapiddetection and identi® cation of outlier moisture and FOFbobbins during production. Non-contact, at-line moist-ure and FOF measurements directly on the bobbin inmanufacturing would result in improved yields, processmonitoring, and Quality Assurance. In this work weestablish the feasibility of using a portable Near InfraRed(NIR) moisture analyser (Kett KJT100) to measuremoisture directly on nylon tire and carpet yarn bobbinsin manufacturing and demonstrate the feasibility of asubjugate process monitoring measurement of bobbinFOF during spinning. NIR moisture calibrations weredeveloped for numerous tire and carpet products (diå er-ent yarn type, size, ® nish, etc.) at three locations(laboratory, lag area, spinning). The NIR methodsuccessfully monitored moisture diå erences between bob-bins in the laboratory and manufacturing areas. Inspinning, a strong correlation was observed betweenNIR bobbin moisture and FOF. Subjugate FOF calibra-tions for tire and carpet products were developed inspinning, and excellent agreement was observed betweenthe NIR and reference FOF results. Of critical im-portance to manufacturing was the rapid at-line identi-® cation of several ` outlier’ bobbins, preventing theircontamination of downstream processes. The impacts ofyarn parameters, measurement location, and environ-mental and operational conditions on the NIR resultswere slight.

Novel applications of Raman spectroscopy to pro-cess monitoring and materials’ characterization

Brian J. Marquardt, CPAC, Center for Process Analytical Ch,FJ-20, Seattle, WA 98195, USA

This presentation will focus on the use of Raman spectro-scopy for the analysis of solid samples (powders, slurries,etc.) with emphasis on on-line process analysis applica-tions. A novel high-precision Raman probe for on-lineprocess analysis will be described. The unique design ofthe Raman probe provides enhancements in measure-ment precision by increasing the reproducibility andaccuracy of optical sampling of high solid contentsamples. The probe has been proven an eå ective sam-pling interface for the analysis of powders, suspensions,slurries, particles and solids. The ease of use of theRaman probe and the increased sampling precision haslead to its use in various proof of concept and on-lineprocess analytical applications. These applications in-clude dry powder± powder mixing eæ ciency, coating


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thickness measurements, solvent drying analysis, reactionmonitoring and various other analytical processes. In thispresentation I will discuss the physical and optical designof the Raman probe and demonstrate its applicability asan on-line sampling tool.

Quantitative in-line measurements of paper coat-ings by near infrared

Jon G. Goode1, Qian Wang1, Angela Schmidt2 and HelmutWeiler2, 1Bruker Optics, Applications, 19 Fortune Drive, Bill-erica, MA 01821-3923, USA, 2Bruker Optik, Near InfraredApplications, Rudolf-Plank-Strasse 23, Ettlingen D-76725,Germany

Leading manufacturers in the paper industry are lookingfor in-line methods to determine the physical properties,perform component analysis and determine appliedcoating weights in paper. The MATRIX-E, an in-lineprocess control FT-NIR spectrometer, makes possible anon-contact diå use re¯ ectance measurement with a largesampling area. We will discuss an on-line paper applica-tion to determine the silicone coat weights on label-stocks. This is currently performed in the laboratory bya time-consuming X-ray ¯ uorescence method. The aimwas to achieve an absolute value for the deviation fromthe target of 1 g/m2 during continuous paper productionat velocities of approximately 400 m/min. Concentrationsof silicone between 0 and 2 g/m2 on various papersubstrates were included in a quantitative model andin uences from the uncoated paper type due to supplier,colour, opacity, area densities, precoatings as well asdiå erent compounds of the silicone agent were investi-gated. It was found that all these factors could beincluded in a single PLS model. The fact that elementalsilicone is present in clay-coated papers was found to beof no consequence to the measurements with MATRIX-E. Moreover, during in-line installations it was foundthat the variation of the moisture content in the movingpaper due to variable machine velocities as well as there¯ ecting material of the cylinder had to be considered. Itwill be shown that the result of the in-line calibration hasthe same prediction capability as laboratory scale results(root mean square error of cross-validation , RMSECV =0.034 g/m2).

Identi� cation of protein folding subunits by ionmobility-mass spectrometry

Brandon T. Ruotolo, Kent J. Gillig, Earle G. Stone and DavidH. Russell, Texas A&M University, Department of Chemistry,PO Box 30012, College Station, TX 77842-3012, USA

There have been several theories proposed as to themechanism of folding processes in proteins. Among them,the idea that certain peptides segments of a proteinexhibit intrinsic stability and contain the site(s) of helixnucleation within the protein, i.e. autonomous foldingsubunits, has been proposed and observed in a fewisolated cases. However, with the introduction of rela-tively new methods for probing the conformation ofbiological molecules in the gas phase, such as ion mobilityspectrometry, additional information can be obtained onthe validity of this hypothesis. Proof-of-concept experi-

ments have been performed on tryptically digestedproteins, which are then screened by MALDI-IM-TOFMS. For example, a peptide signal from a tryptic digestof horse heart myoglobin was observed to deviate bymore than 10% in total drift time from the other peptidesin the IM-MS map. The peptide was sequenced usingtandem mass spectrometry and identi® ed as the majorityof the E helix of solution phase myoglobin. This peptidewas found to exhibit helical structure in simulatedannealing molecular dynamics simulations. This presen-tation will focus on our recent eå orts to probe the abilityof MALDI-IM-TOF MS to screen for the presence ofthese extraordinary peptides. The updated screeningprotocol involves digestion with a number of diå erentproteolytic agents and in diå erent solvent systems inorder to produce a more complete map of the proteinunder investigation. These results will be discussed inlight of insight gained on protein folding mechanisms.

Automatic HPLC method development with intel-ligent peak tracking

Wolf-Dieter Beinert1, Volker Eckert1, Reinhold Spatz1, SergeyGalushko2 and Irina Shishkina3, 1Merck KGaA, Scienti¢c andLaboratory Products, Frankfurter Str 250, Darmstadt D-64271,Germany, 2Institute of Bioorganic Chemistry, National Academyof Sciences, Fine Organic Synthese, Im Wiesengrund 49-b,Muehltal 64367, Ukraine, 3Vsevolod Tanchuck, Institute ofBioorganic Chemistry of National Academy of Sciences, Mur-manskaya 1, Kiev-94, Ukraine

An automated expert system has been developed tosearch for optimum conditions in HPLC. Controlled byan arti® cial intelligence module, the system provides fullyautomated unattended method development in reversed-phase HPLC for mixtures of compounds and can searchfor the optimum concentration of an organic modi® er forisocratic separation, for the best linear and multisegmentgradient pro® les and for the temperature optimum. Forproper peak tracking, the ® rst version of ChromSwordAuto required single standards of the compounds to beseparated. However, often one faces the situation that notfor all compounds to be separated suitable standards areavailable. An example is the analysis of impurities in aproduct, where usually only a few (if any) of theimpurities are available as pure substances. This isusually the case with pharmaceutical quality controlsamples for purity and stability tests. Therefore, Chrom-Sword Auto has been equipped with intelligent peaktracking capabilities. Applying advanced mathematicalprocedures for peak assignment, it is now possible toperform the optimization procedure with the samplemixture. The number of standards necessary for theoptimization procedure can be reduced to just two,regardless of how many compounds are present in themixture. The objects of the automated optimizationprocedure are: (1) separation of the mixture into amaximum number of peaks (e.g. search for impurities)or separation of target compounds; (2) optimum resolu-tion; and (3) minimum analysis time In this way it ispossible automatically to optimize HPLC separations ofchemical and pharmaceutical samples where only a fewsubstances (e.g. the active compounds) are available as


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standards. ChromSword Auto thus makes possible dra-matic timesavings.

The virtual assistant—a new HPLC system withexpert knowledge and best performance

Reinhold E. Spatz1, Wolf-Dieter Beinert1, Masahito Ito2,Honori Kaji2 and Richard Jack3, 1Merck KGaA, SLP Chro-matography, Frankfurter Str 250, Darmstadt D-64271, Germany,2Hitachi Instruments Ltd, Biosystems, Hitachi-Naka 312-8504,Japan, 3Hitachi Instruments, Inc., 3100 N. 1st Street, San Jose,CA 95134-1942, USA

HPLC is by far the most used instrumental technique inthe analytical laboratory. The instrument speci® cationscontinuously need to be adapted to new analyticalrequirements such as diå erent column dimensions, highsample capacity, fast sample throughput, latest govern-mental regulations or state-of-the-ar t informationmanagement. The paper describes the concept of thenewly developed modular LaChrom Elite HPLCinstrument system. It is suitable as well for standardHPLC applications as well for special applicationsegments in HPLC, like semi-micro HPLC or high-throughput separations with monolithic stationaryphases. The same system can be modi® ed by the userwithout compromising speci® cations for the application® eld in target. This ¯ exibility is due to a new hardwareconcept and due to integrated unique software optionsthat act like a ` virtual assistant ’ in the back-ground todeliver highest expertise whenever necessary for a certainapplication. Typical examples that require optimizedhardware are semi-micro HPLC with 1 mm i.d. columnsor High Throughput HPLC with monolithic columnsand ¯ ow gradients in the 10 ml/min ¯ ow rate range.Typical examples for Virtual Assistance are integratedsoftware modules such as ` ChromSword Auto’ for fullyunattended HPLC method development or ` AutoValida-tion’ for automated Operation and Performance Quali-® cation (OQ, PQ) of the HPLC system for regulatedQuality Control Laboratories. The paper will describethe new hardware and software concept. Many evalua-tion results and typical applications will prove the hightechnical level of this new development.

An automated software approach to analyticalmethod validation

Michael E. Swartz and Patricia A. Fowler, Waters Corporation,Pharmaceutical Marketing Lab, 34 Maple Street, Milford, MA01757-3696, USA

Method validation is a tedious process performed todetermine if an analytical method meets the require-ments for its intended purpose. In the regulated labora-tory, method validation may take many days to performthe necessary analytical tests. Data reduction and thestatistical analysis of results performed can be a very timeconsuming process. There is also a greater possibility ofintroducing error when calculations are performedmanually. With the use of automated software to performthese calculations, method validation is much faster andeasier, with less chance for error. In this presentation, wewill show how an analytical method is validated using

automated software. Chromatographic results are di-rectly accessed from a relational database bypassingmanual intervention. Statistical calculations are per-formed automatically and a report generated showingthe results of the analyses from the Student, Cochran,Dixon, and Fisher Tests. Graphs are generated represent-ing the results of the statistical analysis. In addition, wewill show that the data reduction and statistical calcula-tions necessary to validate the method, complete with thenecessary documentation and report generation, arecompleted in signi® cantly less time.

A collaborative environment for developers ofscienti � c software

Deborah A. Kernan, Victoria Rafalovsky and Ty Abshear, Bio-Rad Laboratories, Informatics Division, 3316 Spring GardenStreet, Philadelphia, PA 19104-2552, USA

Bio-Rad’ s KnowItAllTM Analytical System oå ers anintegrated environment for analytical techniques, such asIR, H-1 and C-13 NMR, UV/Vis, GC, MS, Raman,and NIR. Within this system, chemists can performmultiple tasks within the same interface, using software` plug-ins’ that reside within the main KnowItAll archi-tecture. This design allows the user to easily transferinformation from plug-in to plug-in without having toopen another program. Because of this unique design,new third-party software plug-ins can be quickly andeasily incorporated into the KnowItAll architecture. Thephilosophy of this unique architecture is one that builtupon the ideal of getting solutions to scientists faster andgiving them more options. Thus, through collaborationswith database content providers and third-party softwaredevelopers throughout the world, the system grows andadapts to meet the changing needs of the scienti® ccommunity. For those parties with existing software thatmay ® t within the content of the system, Bio-Rad hasdeveloped a simple Software Developer’ s Kit to convertsoftware into a format that will ` plug into’ and workwithin the KnowItAll system.

Development and validation of analytical softwarein a regulated environment

Kevin Bynum, Gillian Raymond, Lane Gehrlein and PhilipPalermo, Purdue Pharma LP, Pharmaceutical Analysis, 444Saw Mill River Road, Ardsley, NY 10502-2605, USA

The development and validation of an analytical soft-ware package for use in the collection of cGMP compli-ant data will be discussed. The system has been designedto collect data to support drug development, stabilitytesting, and release testing in the Pharmaceutical In-dustry. The software is designed to collect dissolutiondata from a UV probe ® bre optic dissolution system. Thesoftware, written in JAVA and C‡‡, uses an Oracledatabase to ensure data integrity and security. Thedesign features, which make this software ` validatable ’will be discussed. The validation testing and implementa-tion of the system on our corporate network will bediscussed in detail.


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Validation Manager 2.0: new developments incomputer-assiste d assay methods validation

Jean-Marc Roussel1, Michel Righezza2 and Wolf-Dieter Bei-nert3, 1Merck Eurolab S.A.S., 201 Rue Carnot, Fontenay SousBois F-94120, France, 2Antenne Scienti¢que Universitaire deChartres, LBC, 21 Rue De Loigny La Bataille, Chartres F-28000, France, 3Merck KGaA, SLP Chromatography, Frank-furter Str 250, Darmstadt D-64271, Germany

For laboratories willing to meet the requirements ofISO 17025 regulations, analytical assay methods valida-tion is an step important to consider. Recommendationsfor these method validations have been announcedduring the fourth International Conference on Har-monization (ICH 4) and the corresponding sta-tistical calculation procedures are described in the ISOguidelines (i.e. ISO 5725, ISO 8466). Based on theserecommendations, we have developed an assay methodsvalidation software which allows validation planning,calculations and reporting according to the authoritiesrequirements (USP, EP, FDA). Validation step consistsin the study of the method characteristics as describedin the guidelines and uses well-known and eæ cientstatistical tests such as Dixon, Fisher, Student orCochran. Although these statistical tools are frequentlyused in the analytical laboratories, most questions comewhen preparing the validation planning or determiningthe correct calculation con® guration to be used. Inorder to help the analyst, we have implemented in thesoftware ready to use validation templates which in-clude, for diå erent analytical techniques, prede® nedstatistical tests con® dence levels and default number ofanalytical results per validation characteristics, all thesein conformity with the international recommendationsand regulations. A validation document preparationwizard may be used in combination with these tem-plates, in order to prepare the detailed validationplanning in the easiest way. Electronic data securityhas become of the utmost importance, in this respectValidation Manager software proposes to the user aFDA compliant con® guration in which every singleaddition or modi® cation in data or result is fullydocumented and saved in a method logbook. Eachversions of the validation document is saved in a speci® cfolder and cannot be overwritten. Each data modi® ca-tion and calculation creates a new results version,ensuring full data integrity. Moreover, in order to avoidtranscription mistakes, an automatic data recoveryfunction has been developed allowing direct copy ofinformation from the Hitachi D-7000 HSM and SSIEZ-Chrom Elite chromatography data systems. Usinganalytical data from our laboratory, the presentationwill describe the diå erent steps of method validationusing Validation Manager. The next future of thisdevelopment project, including the use of experimentaldesigns for robustness study will also be presented.

Advanced automation interfaces: new toolkits foradding instrument control to a chromatographydata system

Dario Fiore and Kristi McKiney, Scienti¢c Software, Inc., 6612Owens Drive, Pleasanton, CA 94588-3334, USA

One of the greatest limitations of chromatography datasystems is in the area of true (interactive) instrumentcontrol for diå erent types of instruments from disparatemanufacturers. Because instrument manufacturers are inbusiness to sell instruments, many feel it is not in theirbest interest to provide control for competitive instru-ments. Yet this means the user must learn to operatemultiple data systems in a single lab, many of whichmight not be the data system of choice. Hundreds ofman-hours are wasted in learning, maintaining, andvalidating diå erent data systems that essentially do thesame thingÐ data acquisition, analysis and instrumentcontrol. Many companies (both data system users andinstrument manufacturers) are realizing the time andcost bene® ts of using open-architecture data systemswhere instruments of many brands are supported.Although adding control for an analytical instrument isnot a task for the everyday user, new tools are nowavailable that facilitate the interfacing of instrumentsto the EZChrom Elite Chromatography Data System,making it much easier and faster for instrument manu-facturers and large companies to create an instrumentcontrol interface. The Toolkit automation interfaceprovides the tools required for a quali® ed company toadd instrument control, and the new Rapid Controlproduct allows Active X control to be added to the Elitedata system. This paper will describe these tools and giveexamples of their use.

On-line analysis with a liquid sampling-atmos-pheric pressure glow discharge (LS-APGD)

R. Kenneth Marcus and W. C. Davis, Clemson University,Department of Chemistry, 312 Hunter Laboratory, Clemson, SC29634-0001, USA

The successful implementation of any form of detection inmicro uidic (gas or liquid) streams is a matter of both theobtained ® gures of merit as well as the compatibility ofscale regarding the instrumentation and the concurrentcapital costs. In the case of liquid sampling in the realmof capillary liquid chromatography or chip-based separa-tions, the use of a multiple kilowatt plasma whichoperates in the 10 litres/min gas ¯ ow regimes (i.e. aninductively coupled plasma) does not make practicalsense. In this laboratory, we have developed a liquidsampling-atmospheri c pressure glow discharge (LS-APGD) optical emission source as a detector for ¯ owinjection or liquid chromatography separations. A lowpower is used though with suæ cient energy to consumethe eluting liquids totally at ¯ ow rates up to 0.5 ml/min.The components of the apparatus are depicted in the® gure. The plasma operates in the abnormal glowdischarge regime at currents of 30± 80 mA and potentialsof up to 2000 V. The LS-APGD is actually created suchthat the liquid sample acts as an electrode, in fact eitheras the anode or cathode. A simple sheath gas is employedin the current embodiment to alleviate pump-derivedpulsation in the liquid ¯ ow when rates of ml injections. Inabsolute terms, this equates to about 10 ng analytes (Na,Mg, Hg, Pb) in a single injection. In this presentation, wewill describe the operating characteristics of the LS-APGD with regards to various powering modes and its


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use as a detector in reverse-phase liquid chromatography.In addition to the metals listed above, the determinationof halogen elements will also be described. Given thecompact size, experimental simplicity and sensitivityobserved to date, it appears that the device should havegreat promise as a detector for capillary LC or chip-basedseparations in the areas of environmental and potablewater systems.

A novel molecular aptamer beacon for real-timeprotein recognition

Jinawei Je¡ery Li1, Weihong Tan1 and Xiaohong Fang2,1University of Florida, Department of Chemistry & TheMcKnight, PO Box 117200, Leigh Hal, Gainesville, FL32611-7200, USA, 2University of Florida, Institute of Chemistry,52 Sanlihe Road, Beijing 100080, P. R. China

One of the most pressing problems facing those attempt-ing to understand the regulation of gene expression andtranslation is the necessity to monitor proteinproduction in a variety of metabolic states. Yet, thereis no easy solution that will either identify or quantitateproteins in real time. Recently molecular beacon basedassays have shown promise for real-time proteindetection. Here we introduce a novel approach toconstruct aptamer beacon for real-time proteinrecognition and quantitation. An aptamer beacon basedon a thrombin-binding aptamer was prepared bylabelling the two ends of the aptamer with a¯ uorophore and a quencher, respectively. The aptamerbeacon combines the signal transduction mechanism ofmolecular beacons and the molecular recognitionspeci® city of aptamers. Signi® cant ¯ uorescent signalchange was observed when aptamer beacon was boundto thrombin, which is attributed to a signi® cantconformational change in aptamer beacon from aloose random coil to a compact unimolecular quad-ruplex. The aptamer beacon recognizes its targetprotein with high speci® city and high sensitivity inhomogeneous solutions. Ratiometric imaging has beenconducted with aptamer beacon labelled with two¯ uorophores, which makes it feasible for proteinquantitation in living specimen. The unique propertiesof the aptamer beacon will enable the development of aclass of protein probes for real-time protein tracing inliving cells and for eæ cient biomedical diagnosis inhomogeneous solutions.

Rapid screening of environmental test wells byheadspace mass spectrometry

Brian G. Rohrback1 and Elizabeth A. Harvey2, 1Infometrix,Inc., PO Box 1528, Woodinville, WA 98072-1528, USA,2Chevron Research, 100 Chevron Way, Richmond, CA 94801-2016, USA

Gas chromatography is the preferred method for char-acterizing and quantitating low molecular weight com-pounds, but the separation process is often slow andoften requires sample pretreatment. By eliminating thechromatograph and applying multivariate techniquesto the bulk detector signal, composition can be deter-mined quickly. Using a headspace sampling system

with direct injection into a mass spectrometer, bothqualitative and quantitative precision are demonstratedfor a variety of environmental test applications. Theshort (3± 5 min) analysis time and the lack of samplepreparation create a fast screening technique for fugi-tive hydrocarbons. Although headspace technologieshave gained some acceptance in the ¯ avour andfragrance industry as a means of evaluating productand ingredient quality, little has been done to assessfeedstocks, products or wastes in the petroleum in-dustry. Data for this study are drawn from a set of200 + test wells positioned at various locations withinthe boundaries of a re® nery. For traditional qualityassessment, classi® cation techniques (such as nearest-neighbour and factor-based groupings) can be used todetermine the contaminant origin even in cases wherethe candidate sources are chemically similar. Withoutresorting to GC separation, the concentration of classesof contaminants can be estimated over a four order-of-magnitude range.

Personal Digital Assistant (PDA) LIMS: the inte-gration of LIMS (Laboratory Information Manage-ment System) and mobile technology

Russ Vranken, Don Kolva, Tom Miller and Christine Paszko,Accelerated Technology Laboratories, Inc., 496 Holly GroveSchool Road, West End, NC 27376-8412, USA

As technology advances, more options become availablefor developers to create products that are both practicaland functional for today’ s mobile generation. It hasbeen proven in the past that automation is a vitalkey to the success of a Laboratory. The handhelddevices of today are smaller and more powerful thanthey have ever been. For example, when Palm, Inc.® rst introduced the Pilot, it had a mere 512 k of RAMwhich was good for the apps that were already integratedinto the device, but limiting for additional applications.The PDAs of today have seemingly unlimited amounts ofRAM (considering a Palm application typically takes upbetween 10 and 60 k RAM) and a plethora of optionsmaking application development for these PDAs limitedonly by the imagination. One area of the LIMS that is inneed of these types of applications is the sample login.With a LIMS application on the PDA, laboratorytechnicians in the ® eld can easily enter the informationabout a collection session, return to the laboratory andsimply ` hotsync’ or upload to the LIMS with thatinformation. Another function that will ease the burdenof sample login will be to have pre-logged samples(samples waiting to be logged into the LIMS) sent tothe PDA during synchronization and the laboratorytechnician in the ® eld can simply enter the pertinentinformation related to the samples downloaded andsynchronize back to the LIMS, automatically loggingthe samples in. In conclusion, the evolution of PDAtechnology and LIMS will allow laboratories to becomemore eæ cient in all aspects of sample and data manage-ment. As PDAs continue to advance, it is foreseeable thata full-featured LIMS could be implemented on thesedevices allowing laboratories to become even moremobile and eæ cient.


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Using software development methods for labora-tory information management systems imple-mentations

Nicholas J. Arnold, Thermo LabSystems, 1 St George’s Court,Altrincham WA14 5TP, UK

As with many computer and other systems projects, theimplementation of a Laboratory Information Manage-ment System (LIMS) can be a major undertaking for anorganization. The success of a LIMS implementationproject will depend on a number of factors includingthe technology employed, the composition of the projectteam and the involvement and buy in of the user com-munity. Over the years, many software developmentmethods have been designed or evolved, some of whichcan be used by project teams to facilitate their LIMSimplementations. This paper will review a number ofsoftware development and project management methodsthat have been adapted for use in LIMS implementa-tion projects. Particular emphasis will be placed on theuse of the Dynamic Systems Development Method(DSDM), which is a Rapid Application Development(RAD)-based approach, several features of which havebeen employed by project teams on LIMS implementa-tions in a variety of environments. Particular DSDMelements, which have been most successfully taken upand applied, include: user involvement, the use offacilitated workshops, high-level requirements, 80/20thinking and prototyping The bene® ts of using suchapproaches will be discussed, along with the pitfalls. Inaddition, typical risks pertaining to LIMS projects, alongwith methods for assessing and mitigating them, will bepresented.

The role of di´ usive sampling in the ambient airmonitoring in Europe

Richard H. Brown, Health and Safety Executive, Broad Lane,She¤eld S3 7HQ, UK

Most early applications of diå usive sampling were formonitoring in industrial environments, where its sim-plicity and user friendliness gave it a positive advantageover sampling pumps. The EC Chemical Agents Direc-tive, 98/24/EC, requires an assessment of a worker’ spotential exposure to chemicals as part of an assessmentof any risk to his/her health and safety. This in turngenerally requires measurements to be taken that needto be as cost-eå ective as possible to minimize anyburden to industry. Thus, diå usive sampling is an idealcandidate. Similarly, the application of diå usive sam-pling techniques to ambient air has been given addedimpetus by European legislation, in this case theAmbient Air Directive 96/62/EC and its Daughters.These also require an assessment of air quality, byrepresentative measurements, surveys or other means;it being assumed that measurements near the limitvalues signify a signi® cant risk to the population orthe environment. Depending on the anticipated con-centration levels, diå erent regimes and quality objec-tives for measurements are speci® ed, and it isanticipated that diå usive sampling will be especiallyuseful for ` indicative’ measurements. The task of devel-oping appropriate standards for workplace air quality

measurements within the European Community hasbeen carried forward by working groups of CENTechnical Committee TC 137. It took the view thatair quality assessment standards should take the form ofperformance requirements rather than prescribedmethods. This approach has the advantage of allowingany method to be used that meets these requirementswithout sti¯ ing innovation and development. Thus,CEN/TC137/WG2 has developed a hierarchy of stan-dards with a general performance requirements docu-ment, EN 482, at the top and a series of specializedstandards (e.g. EN 838 for diå usive sampling) underthis umbrella. The parallel CEN committee for devel-oping appropriate standards for ambient air qualitymeasurements is TC 264. The primary task of thisTC is to evaluate methods and recommend referencemethods where these are not already prescribed inDirectives. In the speci® c case of diå usive samplers,performance requirements standards (prEN 13528-1and -2) have been developed by CEN/TC264/WG11.These draft standards are now being prepared for FinalVote and should be publicly available early in 2002.

Advances in Membrane Extraction with SorbentInterface (MESI) for on-site continuous air qualitymonitoring

Janusz Pawliszyn, University of Waterloo, Department ofChemistry, 200 University Avenue West, Waterloo, OntarioN2L 3G1, Canada

The ultimate goal of chemist is to perform analysis at aplace where a sample is located rather than moving thesample to the laboratory, as is common practice in manycases at present. This approach eliminates errors and thetime associated with sample transport and storage and itwould result in more accurate, precise and faster analy-tical data. In addition to portability, two other importantfeatures of ideal ® eld sample preparation technique arethe elimination of solvent use and integration with asampling step. These requirements are met by techniquesusing polymer technologies. Membrane Extraction witha Sorbent Interface (MESI) uses a polymeric membranewhich is in contact with a sample ® tted directly into acarrier gas line of a gas chromatograph equipped with asorbent trap. Analytes partition into the polymeric phaseof the membrane and after diå usion through the mem-brane are carried by the gas to the sorbent trap.Periodically the concentrated analytes are delivered ontothe front of the column by a thermal pulse. MESI is adynamic system where the rate of analyte intake isdependent on both the diå usion coeæ cients of analytesin the membrane material and the membrane/samplematrix distribution constant. Similar as SPME, MESIcan be used for both spot- and time-averaged monitoring.Both SPME and MESI techniques integrate samplingwith sample preparation and sample introduction toanalytical instrument into a simple procedure. They aresolvent-free techniques that not only facilitate convenient® eld sample preparation, but also rapid sample intro-duction resulting in fast chromatographic separations. InSPME, mechanical movement of the ® bre is necessary;however, sampling and sample introduction steps areseparated in space allowing one instrument to analyse


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large number of ® bres. MESI on the other hand requiresdedicated, permanently attached instrument to one orseveral membrane/sorbent systems, but it eliminates theneed for mechanical movement and therefore reducespossibilities of failure. Fundamental aspects of TWAsampling by both techniques will be emphasized in thetalk and their performance will be compared.

Di´ usion-based air sampling with solid-phasemicro-extractio n for indoor and ambient air

Jacek Koziel1, Jarett Spinhirne2, Darren Williams2 and DavidParker2, 1Texas A&M University, Texas Agricultural Experi-ment St, 6500 W. Amarillo Blvd, Amarillo, TX 79106-1796,USA, 2West Texas A&M University, Canyon, TX, USA

Solid Phase Micro-Extraction (SPME) combines sam-pling and preconcentration. SPME uses extracting poly-mer coated on a fused silica ® bre that is movable inside/outside of SPME needle assembly. Such a SPME con® g-uration can be easily adapted to on-site air sampling andcoupled with direct transfer of extracted analytes into astandard (or portable) gas chromatography (GC) system.Complete thermal desorption of analytes in a GC injectormakes SPME ® bres reusable. In the last decade, thetheory of sampling with SPME was developed andapplied to environmental samples including airbornevolatile organic compounds (VOCs). These extractionsare controlled by diå usion of VOCs onto the SPMEcoating, particularly when the SPME coating is signi® -cantly far from saturation. Diå usion-based calibrationsare used for (1) time-weighted average (TWA) and (2)rapid sampling. For TWA sampling, SPME ® bre coatingremains retracted and analytes diå use to ® bre coatingfrom the needle opening. The knowledge of gas-phasemolecular diå usion coeæ cients (Dg), time (t), diå usionpath (Z), needle opening area (A) and mass extracted (n)allows for VOC determination in air. SPME in the TWAsampling mode is applicable to long-term sampling. Forrapid (spot) sampling, typically lasting < 1 min, VOCsdiå use through the boundary layer around the ® brecoating that is exposed to forced air ¯ ow. VOC determi-nation is based on the knowledge of Dg, t, n, length (L)and radius (b) of the SPME ® bre, and the thickness of theboundary layer. This presentation will entail an overviewof rapid and TWA SPME methods for determination ofVOCs in air. New developments in rapid and long-termsampling will be illustrated with ® eld data from indoorair surveys and agricultural odorous gases. Advantagesand challenges associated with ® eld air analysis withSPME will also be discussed.

Selenium speciation by liquid chromatography:particle beam/hollow cathode optical emissionspectroscopy: monitoring of carbon and hydrogenemission in selenoamino acids

Fuxia Jin and R. Kenneth Marcus, Clemson University,Department of Chemistry, 312 Hunter Laboratory, Clemson, SC29634-0001, USA

As an essential trace mineral in the human body,selenium (Se) participates in anti-oxidative, cancer che-mopreventative and immune-improving processes. Since

the biochemical function of Se is highly species-depen-dent, the speciation of Se compounds rather than total Sedetermination is extremely important. Ion exchange andion pair reverse-phase HPLC mainly have been appliedto the separation of Se species. An ion pair reverse-phaseHPLC method for the separation of selenomethionine,selenoethionine and selenocystine was optimized to havea mobile phase composition of 95% water± 5% methanolcontaining 0.1% TFA [1]. A diversity of methods wasemployed for the detection of Se content, which includeselectrochemistry, UV detection, ¯ uorometry, atomicspectroscopy and mass spectrometry. We have success-fully developed a new liquid chromatography methodfor the separation and speciation of organic and inor-ganic Se compounds via liquid chromatography /particlebeam hollow cathode optical emission spectroscopy sys-tem (LC/PB-HCOES) [2]. A detailed systematic studyof selenoamino acids separation on four commercialreverse-phase C18 columns by reverse-phase chroma-tography has been performed. The baseline resolvedseparation of selenoamino acids by reverse-phase liquidchromatography with a high methanol content up to15% in the mobile phase without the presence of any ionpair agents has been achieved. This greatly facilitates thesample preparation procedure and favourably enhancesthe feasibility of interfacing liquid chromatography toparticle beam system. The successful separation of sele-noamino acids with a mobile phase of only water andmethanol makes it possible to identify the selenoaminoacids through the analysis of carbon and hydrogenelements. The carbon and hydrogen optical emission inthe selenoamino acids was monitored for the LC eluentsafter separation. The eå ects of solvent composition andglow discharge conditions on spectroscopic backgroundwere studied. The analytical response curve for inte-grated C and H emission intensities as a function of theconcentration was obtained.

1. Puskel, E., Mester, Z. and Fodor, P. J. Analytical and AtomicSpectrometry, 14 (1999), 973.

2. Davis, W. C., Jin, F., Dempster, M. A., Robichaud, J. L.. andMarcus, R. K. J. Analytical and Atomic Spectrometry (in press).

Method 1631: mercury holding time study

Mark L. Bruce, Scott Irwin and Patrick Omeara, Severn TrentServices, STL North Canton, 4101 Shu¡el Drive NW, NorthCanton, OH, 44720-6935, USA

US EPA Method 1631C is required or recommended fortotal mercury analysis in many types of water matrices inthe low parts per trillion concentration range. Currently,the method requires that the water sample be preservedwith either HCl or BrCl within 48 h of sample collection.Preservation in the ® eld involves the shipment of hazar-dous chemicals and additional manipulations that in-crease the risk of accidental contamination. Thus,preservation at the laboratory is preferred. To accom-plish laboratory preservation currently necessitates over-night shipment on the day of collection and day of receiptpreservation at the receiving laboratory. This rapidaction in the ® eld and in the laboratory is possible mostof the time but there are numerous instances where thisrapid sample handling is impractical, expensive and, in


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some cases, impossible. Lengthening the unpreservedholding time would facilitate more eæ cient samplehandling both in the ® eld and at the laboratory. Aholding time study was undertaken to demonstrate thattotal mercury content in water sample is stable for muchlonger than 48 h. Four diå erent types of water samples(reagent water, freshwater pond, industrial e¥uent andpublicly owned treatment works e¥uent) with concen-trations between 1 and 5 ng/l were studied over 35 days.Aliquots of each sample were held unpreserved for 7, 14,21, 28 and 35 days. The samples were then analysed byMethod 1631. Statistical comparisons of the linear least-squares regression line of each sample data set indicatesthere was no signi® cant change in concentration duringthe 35 days of the study. Thus, the unpreserved sampleholding time for total mercury as measured by Method1631 should be extended to at least 28 days provided thesample is oxidized in the original sample collectioncontainer. The e¥uent from a publicly owned treatmentworks shows stability of the unpreserved sample over a35-day test period, the variation being not signi® cantlydiå erent from zero at 0.01 ng/l/day.

The direct mercury analysis: a revolutionary ap-proach in mercury analysis

Mikhail Mensh, Milestone, Inc., 160B Shelton Road, Monroe,CT 06468-2545, USA

For many years, the bottleneck of mercury analysis wassample preparation. To perform analysis of a sampleusing one of the traditional methods such as cold vapour,the sample has to be properly collected, stored andprocessed (digested and chemically treated). Those stepsare the most time, labour and money consuming. The® nal analysis step takes only a few minutes per sample.Direct analysis of mercury using a Direct MercuryAnalyzer (DMA-80) aå ords many bene® ts. Eliminatingwet chemistry greatly reduces waste generation, systema-tic errors and technician exposure due to volatilization ofmercury during sample handling. Direct analysis typi-cally provides an answer in approximately 5 min after asample has been introduced into the instrument. A widedynamic range, from 0.5 to 600 ng total mercury in asample, is easily accommodated. Con® guring the instru-ment for deployment in the ® eld can reduce systematicerrors due to improper sample storage. Direct mercuryanalysis in the ® eld, near remediation and clean-upactivities can result in signi® cant improvements in theaccuracy of sample data. On-site analysis can signi® -cantly impact the dynamics, as well as the economics, ofremediation activities by providing timely informationfor site management. DMA-80 uses the integrated se-quence of Thermal Decomposition, Catalytic Conver-sion, Amalgamation and Atomic AbsorptionSpectrophotometry . It is described in EPA Method7473 and is validated for laboratory and ® eld analysis.

Validation of EPA Draft Method 3200: mercuryspeciation by selective solvent extraction and aciddigestion

Mizanur Rahman1, Ye Han1, Helen M. Boylan1, Sejal Shah2

and H. M. Kingston3, 1Duquesne University, Chemistry Depart-

ment, 600 Forbes Avenue, Pittsburgh, PA 15282-0001, USA,2Duquesne University, Department of Chemistry and Biochem-istry, 308 Mellon Hall, Pittsburgh, PA 15282-0001, USA,3Duquesne University, 1225 E. Arques Avenue, Sunnyvale, CA94085-4701, USA

The need for speciation analysis, i.e. the determinationof speci® c chemical forms of an element, arises from thenecessity of determining the concentration of thosespecies, which are characterized by the highest toxicity,environmental mobility and tendency to accumulate inliving systems. Mercury species can undergo a variety oftransformations in the environment owing to the inter-conversion and degradation processes, especially forcomplex solid matrices such as soils, sediments andbiological tissues. One of the most important processesis methylation. Investigations showed that such methy-lation process mainly observed in samples rich inorganic matter and high in inorganic mercury. Thistransformation can also occur during extraction andacid digestion processes. The extraction of mercuryand mercury compounds from environmental , biologicaland botanical sample is very complex. The extractionshould be performed in such a way that the analyte isseparated from the interfering matrix without loss,contamination or change of the speciation, and withminimum or no interference. A draft Method 3200,Mercury Species by Selective Solvent Extraction andAcid Digestion, has previously been developed to ex-tract selectively ` mobile and toxic’ species such asalkylmercury, soluble inorganic mercury from soils andsediments. This study mainly focuses on the validationof Method 3200 using standard soil samples. High-performance liquid chromatography (HPLC) is coupledwith the most sensitive elemental detection methodinductively coupled plasma mass spectrometry (ICP-MS) to perform mercury speciation studies. Method7473, a rapid technique for the determination of totalmercury in environmental and biological samples, isused to compare results obtained form HPLC-ICP-MS. Interlaboratory data demonstrating method eå ec-tiveness are also presented and evaluated.

Ageing e´ ects on mercury speciation in soilsamples

Manuel Valiente and Xavier Gaona, Universitat Autonoma deBarcelona, Department de Quimica, Edi¢ci CN, Bellaterra,Barcelona E-08193, Spain

Because of the diå erent bioavailability and toxicity ofinorganic and organo-metallic species of mercury thatcan be present in metal-contaminated sites, a corre-sponding speciation procedure for the proper determineof the content of the diå erent metal species must beapplied. For soil samples, the most accepted procedureincludes the solvent extraction of organo-mercury spe-cies such as methyl-mercury (MeHg) and phenyl-mer-cury (PhHg) among others. Besides volatilization, one ofthe key process that takes place in this system is thedegradation of organo-mercury species with time. Age-ing eå ect will account for both the corresponding lossand transformation of organo-mercury species. Thepresent work is dedicated to ascertain some steps of


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the sample treatment and analytical procedure thatin uences this ageing eå ect. For organo-mercury deter-mination, we used the capillary electrophoresis tech-nique provided with a UV detector. We identi® ed threesteps that in¯ uence the ageing eå ect: sampling, prep-aration and conservation of standards solutions, andconditioning and storing of the soil sample extract.The results, obtained by monitoring MeHg and PhHgin soil samples, show a clear diå erence on the behaviourof both species. Thus, the ageing eå ect for MeHg isdiå erent than for PhHg in the sampling procedure (useor absence of sample freezing). On the other hand, theuse of cysteine as a complexing agent in both sampletreatment and preparation of standard solutions in¯ u-ences the ageing eå ect in a diå erent manner. Thus, thestandard calibration curve has a better repetitivity whencysteine is added to the organo-mercury standard justbefore the CE measurement. The ageing eå ect is alsoobserved on the resolution of the CE what is attributedto cysteine degradation. The illustration of the resultswill follow the presentation of the experimental method-ology employed in this study.

The application of a new separation technology tothe conditioning of wastewater samples for on-line analysers

David Gri¤ths, Filtra Ltd, Biomedical BuildingöFL1000,Australian Technology Park, Ste G11, Eveleigh, NSW 1430,Australia

The use of sophisticated on-line analysers in wastewatertreatment plants can lead to increased eæ ciency andimproved environmental outcomes. Unfortunately, thediæ culties of sampling the process liquors have inhibitedthe use of such analysers in the wastewater and otherindustries. In particular, the need to separate out a rangeof often sticky, ® brous and generally diæ cult solidswithout using maintenance-intensive equipment has beenan issue for the industry. Filtra Ltd, an Australian start-up company, has developed a new solids/liquid separa-tion technology that has particular application to thepretreatment or conditioning of wastewater samples forsophisticated on-line process analysers. The technologyprovides a reliable sample that can be piped signi® cantdistances with minimal sample degradation before an-alysis. Unlike traditional conditioning devices, the newtechnology is intrinsically non-blocking and has minimalmaintenance requirements. Case histories from both pilotplant and operational facilities are presented. The appli-cation of the technology is demonstrated in a variety ofsituations including fast-loop systems and in multiplexedarrangements where multiple sample sources are ana-lysed using a common analyser.

NoteBookMaker is a completely electronic, totallysecure laboratory notebook

Lance Boynton and Stephen Arpie, NoteBookMaker, PO Box5585, Hamden, CT 06518-0585, USA

NoteBookMakerTM is a completely electronic, totallysecure laboratory notebook. In the pharmaceutical in-dustry, the ability to control and easily access securely

all information pertinent to a speci® c project or a seriesof projects is essential. The capability of performing aquick search can greatly increase eæ ciency and there-fore reduce the cost of a laboratory. In the currentpaper format, to search for relevant information, ascientist must locate the correct serialized notebookand then manually ® nd the correct reference in thatpublication. With NoteBookMaker , a simple electronicsearch can ® nd all the related topics in a matter ofseconds. Even with the easy-to-use graphical user, inter-face security is still held at a premium. In fact,NoteBookMake r meets all the requirements for anElectronic Notebook established by CENSA (Collabora-tive Electronic Notebook Systems Association), a con-sortium group that includes individuals from severaldisciplines including end users, government and indus-try. NoteBookMaker creates legally defensible data andis compliant with 21 CFR Part 11. The ® rst realalternative to paper-based systems, it is extremely easyto use and a breeze for IT professionals to manage.Pages are created on demand and look and print likegenuine pieces of paper. A program that enhances thecommunication between chemists and administratorswhile maintaining the highest level of customizablesecurity is fundamental in today’ s competitive market-place.

Rapid on-site analysis of environmental soilsamples in a mobile laboratory using the pressur-ized solvent extraction technique

Al Kaziunas1, Rolf Schlake1, Mark Mathews2 and MichaelWinslow2, 1Applied Separations, 930 W. Hamilton Street,Allentown, PA 18101-1114, USA, 2KB Labs, 6821 SW ArcherRoad, Gainesville, FL 32608-4720, USA

The need for rapid analyses of environmental soil samplesrequires mobile laboratories to extract and analysesamples on site. Unfortunately, the goal of providinganalytical results in the same day is hindered by timeconsuming, traditional solvent extraction techniques.This paper investigates the use of the pressurized solventextraction technique (EPA Method 3545) to reducethe time required to extract soil samples in a mobilelaboratory. Soil samples were extracted on a compactpressurized solvent extraction system and analysedfor organochlorine pesticides, PCBs and PAHs usingSW846 methods 3031A, 8082, and 8100. Results werecompared with typical oå -site laboratory analyses. Inaddition, a calculation of timesavings and reduction ofsolvent usage is presented.

Portable environmental chemical sampler

Jim Smith and Ron Skinner, Prince Edward Island FoodTechnology Centre, PO Box 2000, Charlottetown, PE C1A7N8, Canada

A prototype portable environmental chemical samplerusing Solid-Phase Micro-Extraction (SPME) has beenmanufactured and evaluated for environmental applica-tions. Its small size and the stability of sampled envir-onmental chemicals on the SPME ® bres makes thedevice suitable for many environmental applications


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including the sampling of pesticide run-oå into rivers,pesticide spray drift and e¥uent monitoring. A carouselof SMPE ® bres is contained within the sampler andeach ® bre is exposed to the environment to be sampled.The sampling sequence can be initiated by varioussensors, cellphone modem or preprogrammed into thesampler microprocessor. A typical sampler applicationwould be to sample river water using a carousel of 48SPME ® bres over a 48 h with each ® bre being exposedfor 1 h. A separate sentinel ® bre in the sampler isexposed for the whole 48 h. Once the environmentalchemicals are adsorbed onto the ® bres, they are verystable, unlike liquid samples taken in bottles. There istherefore no urgency to return the samples to thelaboratory for analysis. When the SPME ® bres arereturned to the laboratory for analysis, the sentinel ® breis analysed ® rst. If it is clean, the carousel does not needto be analysed, saving on analysis costs. If the sentinel® bre is positive for the contaminant of interest, thecarousel of ® bres is placed into the modi® ed GCautosampler and the ® bres are sampled automatically.Field trial evaluations have been conducted and will bepresented in detail.

Process control through real-time plasma moni-toring for endpoint, fault and state-of-health

Pamela P. Ward, Peak Sensor Systems, Research and Develop-ment, 6207 Pan American NE, Albuquerque, NM 87109-3425,USA

Plasma processes used for production purposes are dy-namic environments in which minute changes in plasmachemistry can have devastating eå ects on process out-come. In today’ s competitive environment, manufacturesof semiconductors, printed circuit board or other highdollar products in which plasma is a critical productionstep can not tolerate processes that are not controlled.This paper addresses a real-time, full-spectrum opticalplasma-monitoring technique capable of monitoring allplasma processes, recipes and products for endpoint,occurrence of faults and the overall general state of healthof the plasma environment. Data will be shown that willdemonstrate the increase of throughput, yield and overallquality of products monitored via this technique.Through the application of the ProPak in the semicon-ductor industry in a three year study, results haveindicated that the end user can expect increases tothroughput as high as 35%, yield increases as high as12%, and better device performance and clock speeds.Furthermore, a list of captured fault conditions that haveprevented the loss of millions of dollars of products will beshared along with a methodology to detect and preventany plasma fault that has a distinguishable opticalindicator. In this discussion, the operation of the ProPakplasma monitor on AMAT, LAM and TEL reactors, andthe overall bene® t to the semiconductor industry will beshown. Data from real-world production environmentswill be shared to demonstrate the bene® t of plasmamonitoring. Advancements in automatic process controland sophisticated communication techniques will bediscussed.

High-throughput screening of polyole� ns madeusing single site catalysts

Judy Arroyave1 and Wei Sen Wong2, 1Albemarle, R&D, 8000GSRI Road, Box 14799, Baton Rouge, LA 70820-7403, USA,2Albemarle, PO Box 14799, Baton Rouge, LA 70898-4799, USA

As a leading manufacturer of methylaluminoxanes(MAOs), metallocene catalysts and other catalystscomponents, Albemarle Corporation has a key interestin characterizing their performance in a wide varietyof polymerization reactions. To facilitate this research,Albemarle has developed a high-throughput screeningapproach to incorporate the polymerization reactions,isolation and characterization of these product materials.As part of this eå ort, it became necessary to developa rapid GPC technique that would provide accuratedata and faster analysis time while maintaining dataintegrity. This presentation will give an overview ofAlbemarle’ s high-throughpu t screening programmerelated to catalyst performance. It describes a GPCtechnique that allows the screening of these reactions,determining critical information such as weight-averagemolecular weight, intrinsic viscosity and radius of gyra-tion. Laboratory automation along with a uniquecombination of solvent and specialized columns haveyielded very favourable analysis conditions whilemaintaining an excellent degree of accuracy andreproducibility.

Combinatorial screening of polymer librariesusing rapid chromatographic and related tech-

niques

Miroslav Petro, Symyx Technologies, Inc., 3100 Central Expy,Santa Clara, CA 95051-0801, USA

The combinatorial chemistry approaches supportedby rapid automated analytical techniques lead to asigni® cant acceleration of discovery process in severaldiå erent areas, including material polymer science.At Symyx, we have developed a variety of techniquesand approaches based on interaction, separation,molecular recognition and detection of macromoleculesin solution. Those techniques have been successfullyapplied to a number of diå erent material discoveryprojects either to identify or to characterize rapidly thebest polymer candidates from our libraries of materials.The presentation will give an overview of methodologiesfor rapid determination or assessment of the molecularsize, molecular architecture, chemical composition,multiple distribution pro® les of molecular characteristics,interaction and reaction capabilities of macromoleculesat a speed that is satisfactory to match the speed ofparallel polymer synthesis. The techniques are beingapplied for a large diversity of polymers ranging fromthose soluble only in organic solvents at high tempera-tures to those requiring aqueous or ionic environment.In addition, several aspects related to development ofthese methodologies and their application for identi® ca-tion of hits in our polymer discovery programmes will bediscussed.


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Automated methods for characterizin g equilib-rium properties and non-equilibrium propertiesin polymer solutions

Wayne F. Reed, Tulane University, Physics Department, SternHall, New Orleans, LA 70118, USA

Automatic, continuous mixing techniques (ACM) can beused to characterize a vast number of equilibrium andnon-equilibrium characteristics of polymer solutions.Using ACM with combinations of light scattering,viscometric, ultraviolet absorption and refractometricdetectors, without using any chromatographi c col-umns, examples of the following will be given. (1) Auto-matic on-line monitoring of polymerization reactions(ACOMP), which furnishes molecular mass, monomerconversion and other characteristics. Applications ofACOMP to free radical, controlled radical and stepgrowth reactions will be discussed. Recently adapted tocopolymerization reactions, ACOMP allows simul-taneous, on-line determination of copolymer compositiondistributions, sequence lengths, mass distributions andreactivity ratios. (2) Determination of kinetics andmechanisms involved in polymer degradation, which alsoyields information on polymer structure, such as branch-ing and multiple stranding. (3) Monitoring of solutioninstability in the form of aggregation, phase separationand micro-crystallization. ACM is also readily adaptableto monitoring the dissolution of dry polymers. In addi-tion, ACM methods have been adapted to understandcomplex interactions in multicomponent systems, such asthose containing polymer and surfactants, and polyelec-trolytes and salts. Finally, new instrumentation formassive parallel testing and high-throughput screeningwill be mentioned.

Rapid screening of drugs for forensic toxicologyusing retention time locking with gas chromato-graphy/mass spectrometry

Michael Szelewski1, Fran Diamond2 and Paul Miller3, 1AgilentTechnologies, 2850 Centerville Road, Wilmington, DE 19808-1610, USA, 2National Medical Services, 3701 Welsh Road,Willow Grove, PA 19090-2910 , USA, 3National MedicalServices, 2100 Clearwater Drive, Des Plaines, IL, 60018-1918,USA

Identi® cation of drugs in a biological matrix is challen-ging due to interferences and analyte degradation. Atypical analysis involves a library search of all peaksfound in a GC/MS run followed by a manual con® rma-tion by a trained analyst. Retention time locking allowsexact retention times on one instrument or across mul-tiple instruments. These exact retention times can bereproduced after column maintenance, which is a dailyprocedure in many laboratories. A retention time lockeddatabase of 300 drugs and anayltes of interest to forensictoxicologists has been developed. Chromatographicpeaks are searched against the database ® rst for retentiontime and then for target and quali® er ion ratios. A crosscorrelation against the library spectrum is also performedautomatically. Combining retention time with spectralinformation reduces the number of false-positives and -negatives. This rapid screening process for 300 analytes iscompleted in < 1 min after the chromatographic run.

Achieving new levels of reliability in automatedchromatography

James A. Schibler1, Martina Oefelein1 and Andreas Becker2,1Dionex Corporation, 1228 Titan Way, Sunnyvale, CA 94085-4074, USA, 2Dionex Corporation, Via G. Fantoli 16/15, MilanI-29138, Italy

Most modern chromatography laboratories use softwareto automate instrument control and data processing inorder to minimize the operator time required per analy-sis. Although automation can substantially increaseproductivity when everything is working well, mostautomated systems are unable to respond eå ectivelywhen hardware or method problems occur. Commonproblems that a¥ict automated systems include poweroutages, computer-related problems (such as networkcommunication failures or data storage problems), in-strument component failures and inadequate chromato-graphic performance (peak tailing, coelution, poorprecision, etc.). Whether the problem causes the systemto halt prematurely or to continue processing sampleswithout generating usable data, the consequences arecostly: valuable samples and reagents can be wasted,and timeÐ the most precious resource of allÐ is lost. Thispresentation will discuss how a modern chromatographymanagement system intelligently and automaticallyhandles such contingencies, thus providing new levels ofreliability and productivity improvement in automatedchromatography.

Monitoring with MESI

Heather Lord1, Limei Wang1, Rick Morehead2 and JanuszPawliszyn3, 1University of Waterloo, Chemistry, 200 UniversityAvenue West, Waterloo, Ontario N2L 3G1, Canada, 2RestekCorporation, 110 Benner CIR, Bellefonte, PA 16823-8497, USA,3University of Waterloo, 29 Business Park Drive, Branford, CT06405-2967, USA

Membrane Extraction with Sorbent Interface (MESI)technology has been used for continuous monitoring offorensic, environmental and fragrance samples. It hasbeen used for both spot sampling of air concentrations ofchemicals as well as integrative sampling to assess time-weighted average concentration . The feasibility of MESIfor breath sampling was assessed with the analysis ofbreath exhaled while smoking, and with the analysis ofbreath alcohol after consuming an alcoholic beverage.End tidal breath samples were collected and analysed ina sampler designed to trap the last 250 ml of breath.Early breath samples were collected by exhaling into asampling jar. PDMS membrane and a DVB trap and 10-min trapping times were used for all samples with breathsample analysis by GC/FID. Several signi® cant addi-tional peaks were observed in the breath of a smokerduring smoking, as opposed to just before smoking.Ethanol was easily observed in the breath after alcoholconsumption, and the rate of disappearance was mon-itored. Freshly fallen snow was analysed for the presenceof BTEX compounds adsorbed during snow¯ ake forma-tion. Peaks corresponding to benzene, toluene and xy-lenes were observed with signal-to-noise ratios of 3:1 to10:1 by GC/MS analysis from the headspace of 300 ml ofmelted snow. Finally, eucalyptus volatiles were moni-


191

tored from both fresh and day old leaves. All of the majormonoterpene components, as well as some of the lightersesqui-terpenes and green leaf volatiles, were observed.Variation of eucalyptus volatiles could be monitoredrelative to both maturity and senescence states of theleaves.

Improving Raman shift measurement using auto-matic simultaneous Raman and laser calibration

Jun Zhao and Mike M. Carrabba, Chromex, Inc., 2705 PanAmerican Fwy NE, Albuquerque, NM 87107-1630

Accurate and precise measurement of Raman shift valueis of paramount importance to Raman spectroscopy,particularly for demanding in-line monitoring applica-tions. Since the Raman abscissa is a shift relative to thelaser frequency, both the Raman and the laser frequencymust be known. The wide spread use of compactsemiconductor lasers and solid state lasers has com-pounded the problem due to the frequency instabilityof the excitation source. Unfortunately, conventionaldisperse Raman spectrometers are not equipped tocharacterize the excitation source. A new generation ofdisperse Raman instrument was developed to cope withthis diæ culty by simultaneously measuring the Ramanand the laser spectra and calibrating them by the meansof accurately known atomic emission lines. A single CCDdetector attached to a high-quality imaging spectrographcollected all the data. In this study, a 532-nm frequency-doubled Nd:YAG laser from a well-known vendor wasused and its frequency monitored. The inherent fre-quency instability of the laser caused large systematicerrors in Raman shift if it was uncounted for. The newinstrument characterized the laser frequency for eachRaman measurement and yielded consistent results.Precision was improved by an order of magnitude to> 0.05 cm¡1. Such reliable measurement will bene® tRaman spectroscopy for industrial process applications.

Automated interpretation of multivariate instru-ment data

Marlana Blackburn, Scott Ramos and Brian Rohrback, Infome-trix, Inc., PO Box 1528, Woodinville, WA 98072-1528, USA

Several commercial packages for chemometric dataanalysis are currently available, including Pirouette.InStep, a free companion programme, is a custom clientcommunicating directly with Pirouette. It automatesmultivariate prediction and allows the user to customizeeasily an analysis scheme and report format. In itsprocessing mode, InStep periodically checks a watchedfolder for a ® le of speci® ed extension, e.g. a typeproduced by a spectroscopic or chromatographic dataacquisition system. When a new ® le appears, predictionsare made on its data using an InStep method. Tabularresults can be displayed on screen and/or written to a ® le.Control charting of predicted values is also provided. Inits set-up mode, the user can create methods and designreports. An Instep method is a recipe for processing eachsample. The simplest method consists of one model;however, additional processing can be triggered bytesting results of the initial prediction and branching

conditionally. For example, if the predicted Y2 fromModel A is greater than a user supplied value, applyModel B, otherwise apply Model C. The comparisonsand branching can continue inde® nitely.

Optimizing purge-and-trap/mass spectrometrysystems for high performance

Cameron George1 and Philip L. Wylie1,2, 1Agilent Technologies,Technical Support, 91 Blue Ravine Road, Folsom, CA 95630-4720, USA, 2Allen K Analytical Sciences, Five Moore Drive,RTP, NC 27709, USA

The analysis of volatile organic compounds (VOC) invarious matrices is a primary concern for most environ-mental testing laboratories. Recent changes in MSdetector design have resulted in improved sensitivityallowing for potential method improvements. With theseimprovements has come the need for optimizing allsystem parameters, including purge-and-trap conditions,column choice, detector settings and tune values. Thispaper discusses the eå ects of these changes on sensitivity,linearity, analyte resolution and analysis time. An opti-mized set of conditions for the analysis of EPA Method8260B will be considered in detail and all instrumentconditions will be detailed.

Improvements on blood alcohol content determi-nation using autosampler vial headspace sam-pling coupled with gas chromatographic analysis

Charles C. Johnson, Martin Paplewski and John W. Hellgeth,JAS, Inc., 4 Highway Avenue, Ludlow, KY 41016-1663, USA

With the Federal initiatives for highway funding beingdependent upon State compliance with a 0.08% bloodalcohol limit, police and forensic laboratories are underincreased pressure to provide rapid blood alcohol ana-lyses. Classical analyses using conventional headspacesamplers have intrinsic limitations on sample throughput.Previously, a valid method under California Title 17legislation was reported called the Blood Alcohol Report-ing System (BARS). This method demonstrated theaccuracy and precision necessary to comply with theTitle 17 criteria can be accomplished using the headspaceof a sample within a 2-ml Autosampler Vial. Improve-ments to the BARS method, both in the procedure anddata analysis, will be presented.

Chaotic analysis of electrochemica l noise re-sponse of copper

Esteban M. G. Ochoa1, Edgar A. C. Ibanez1 and GuillermoA. V. Coutinho2, 1Instituto Mexicano del Petroleo, ProductionManagement, EJE Central 152, Mexico Distrito Federal,Distrito F 07730, Mexico, 2Instituto Mexicano del Petroleo,Iztapalapa, Mexico

The aim of the present study has been to analyse thecurrent oscillations during the pitting corrosion process ofcopper, joining the information oå ered by traditionaltechniques of electrochemistry (voltammetry), such asreverse-scan polarization, with the analysis of time seriesof the non-linear current oscillations under potentiostaticconditions during the corrosion process. Furthermore, the


192

parameters obtained from the application of the chaostheory allow us to determine whether or not the phenom-enon is of chaotic nature, and to assess the number ofnecessary diå erential equations to describe the system aswell. Besides, we can say that the dynamic changes infunction of diå erent species. Corrosion produced by pitshas diå erent reactions in each step with its own strangeattractor. Recent developments in the theory of lineardynamics have demonstrated the existence of verycomplex solutions to certain very simple diå erentialequations. These solutions exhibit apparently random¯ uctuations. In the case of dissipative systems, the sol-utions curves eventually remain con® ned to a subset ofthe phase space. This subset is known as an attractor.Chaotic behaviour occurs when a stranger exists.

Automation of in vitro drug liberation using � owmethods

Petr Solich1 and Uli Schaefer2, 1Charles University, Faculty ofPharmacy, Analytical Chemistry, Heyrovskeho 1203, HradecKralove, 500 05, Czech Republic, 2Saarland University, Bio-pharmaceutics and Pharmaceuti, Saarbruecken D-66123, Germany

One of the most widely accepted standard for monitoringof semisolid pharmaceutical s is to determine the rate ofrelease of active pharmaceutical ingredients with respectto the time. Recently, according to the recommendationof FDA, the use of the Franz cell system for liberation oftopical preparations is recommended. The studies shouldprovide the multipoint-pro® le testing, which commonlycan be done by HPLC analysis (usually with UVdetection). Although widely used, this method is time-consuming, with a low frequency of sampling. Therefore,a new methodology based on ¯ ow methods FIA and SIAis recommended for testing of in vitro drug liberation fromtopical pharmaceuticals. Flow analytical methods (¯ owinjection analysis (FIA) and sequential injection analysis(SIA)) are predicted for the automation of analyticalprocedures. The interfacing of computer-aided FIA orfully automated SIA with an sensitive ¯ uorescencedetector and Franz cell leads to a fully automatedmonitoring system capable of providing real-time analy-sis and a multipoint liberation pro® le. Examples ofliberation pro® les for a topical dermatological formula-tion containing the active compound salicylic acid areshown. The use of ¯ uorimetric detector increases theselectivity and sensitivity in comparison with the conven-tional UV detection used in HPLC. The recommendedtime for the analysis of the liberation pro® le is 6 h, usingsampling at 0.5, 1, 2, 4 and 6 h after beginning of theprocedure. We have found that the more frequentsamplingÐ every 2± 6 min, which can be done by usingand automated FIA or SIA systemÐ enables one toconstruct a straight line (release rate) after only about3 h instead of the usually used 6 h. Therefore, it can beconcluded that using the FIA procedure with the poss-ibility of analysing 10± 30 samples per hour, the releaserate can be calculated after about half time of the usualmeasurement time. This example shows the attractivityand potentiality of automated ¯ ow methods FIA andSIA for automated monitoring of liberation of activedrug substances from topical applied dermatologicalformulations.

Rapid detection of genetic variations using anautomated capillary electrophoresis system

Dan Zhu1,2, 1Beckman Coulter, Inc., Life Science ResearchDivision, 4300 N. Harbor Blvd, Fullerton, CA 92835-1091,USA, 2Beckman Coulter, Inc., Little Falls Site, 2850 CentervilleRoad, Wilmington, DE 19808-1610, USA

Capillary electrophoresis (CE) has had a major impacton genomics with its signi® cantly increased productivityover slab gel electrophoresis in DNA sequencing. Asincreasingly more DNA sequencing data become avail-able, more studies are required to understand better theassociation between genomic abnormalities and diseasesas well as the signi® cance of the DNA variations amongindividuals. Single base pair mutations, deletions andinsertions of speci® c genes are known to cause speci® cdiseases while single nucleotide polymorphisms (SNPs)may lead to individual diå erences between diseasesusceptibility and response to treatment. We have de-veloped a non-sequencing approach to detection ofmutations/SNPs in a high-throughput and cost-e å ectivemanner using an automated multifunctional CE system.The approach is based on the fact that, under certaintemperature (range), DNA variant with as little as singlebase pair diå erence from a wild-type sequence itself andthe heteroduplexes formed between the variant and thewild-type may migrate at diå erent rates compared withthe wild-type sequence in a denaturing polyacrylamidegel. In this study, we used this approach to detectmutations at codons 12 and 13 of K-ras gene that ismutated in a variety of human cancers. DNA sampleswith all the possible mutation types at the ® rst base sitesof the two codons were tested and all the mutations weredetected without ambiguity. The results demonstratethat our non-sequencing CE-based approach signi® -cantly speeds up mutation/SNP detection processes withaccuracy no less than that of sequencing and primerextension approaches .

An automatic information system and its use involtammetric analysis

Rashida M.-F. Salikhdzhanova1,2, Alexei I. Gorobets1, NataliaJ. Petrova1 and Damir I. Davletchin3, 1Moscow Institute ofRadio Engineering and Electronics, Radio Constructioning,Lenin’s Square 1 80, Moscow 113114, Russia, 2Moscow Instituteof Radio Engineering and Electronics, Analytical Research &Development, Experimental Station E353/3, Wilmington, DE19880-0353, Russia, 3Moscow Textile Academy, ChemistryDepartment, Moscow, Russia

Voltammetric analysis is based on individual methodsfor every object determination that de® ne operatingconditions for the voltammetric instrumentation. Thesemethods give recommendations on the choice of support-ing electrolytes, the electrode system, the potential rangefor the analysed substance and the voltammetric tech-nique to be used. A system of expertise for compiling anautomated data library for voltammetric methods hasbeen presented. This data library is a database system forautomatic retrieval of information that makes use ofselection criteria such as the objects of determinationcriteria such as the objects of determination and analysis,and background. The system can be used for crating


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specialized databases of voltammetric methods for use inenvironmental monitoring, industry and medicine. Datainclude descriptions of some 1000 voltammetric methods.The information system includes a descriptive presenta-tion of methods in complete form as reported in theliterature and in a concise tabulated form. Information,presented in tabular form, is retrieved automatically fromthe library using the following criteria: object of determi-nation, object of analysis, background, indicator elec-trode and type of voltammetric method.

Automation of Karl Fischer methods for the de-termination of water content

Stephanie P. Wilson1, Lynn Jordan1, Terry Roche2 and EliseCorrigan2, 1Zymark Corporation, Zymark Ctr, Hopkinton, MA01748-1668, USA, 2Zymark Ltd, Systems Engineering, 1 Well-¢eld Preston Brook, Runcorn, WA7 3AZ, UK

In 1935, Karl Fischer published information on thedetermination of water content, which has become theglobal standard over many industries. Automation of theKarl Fischer process has been minimal. With the integra-tion of a standard Karl Fisher volumetric titrator, usingdisposable sample holders, and integration with robotics,automation of this labour-intensive task can result inhigher throughput, improved reproducibility, cost sav-ings and promote standardization across industries in-cluding those in the regulatory environment who mustmeet compliance to 21 CFR Part 11. The authors willprovide information on the process of automating theKarl Fischer determination of water content through theuse of current technology and integration of these tech-nologies.

Electronic nose as a quality control tool in packa-ging industry

Tsung Tan1, Nicolas Mignard1, Quitterie Lucas1, HelgeNickel2 and Alexander Stofell2, 1Alpha M.O.S., 20 AvenueDidier Daurat, Toulouse F-31400, France, 2Basell Polyole¢neGmbH, Industriepark Hoechst Build, Frankfurt D-65926,Germany

The consistent manufacture of high-quality PE pellets fordrinking water applications is extremely important forthis industry in general and for Basell GmbH. Tradi-tional methods for taste tests require soaking pellets inwater and conditioning them to carry out the tests. Thenserial dilutions are made until no odour or taste arefound. These tests are time-consuming, require a mini-mum number of sensory panellists and are very repeti-tive. Using these samples, the Fox Electronic Nose wasused to obtain results correlated with these tests. Aftervalidation, the system has been implemented in a QCenvironment. The Fox Electronic Nose has now beenused in QC laboratory for over 2 years. Reliable andobjective results have been obtained in a productionenvironment where the tests are completed four timesfaster than a standard QC sensory panel. These resultswill be presented with emphasis of the reliability of suchinstruments over the long-term, the eå ect of sensorreplacement and also transfer from R&D to QC. Newresults will also be shown on a lower cost, high-through-

put electronic noseÐ GEMINI. The instrument canprovide results every 5 min in a cost-eå ective packagewith high reliability.

Statistical quality control of petrochemical andoleochemical products using the new factory com-patible electronic nose

Quitterie Lucas, Vincent O. Schmitt and Tsung Tan, AlphaM.O.S., 20 Avenue Didier Daurat, Toulouse F-31400,France

Detection of tainted products due to oå -odours and oå -tastes in incoming materials and ® nal products is essentialfor chemical and packaging industries to assess productintegrity and meet customer acceptance especially forfood and pharmaceutical applications. The increasingnumber of controls by factory human panels have ledto the development of a fast, accurate and high-qualityelectronic nose for comparison of manufactured productwith ` gold references’ . A description of a powerful 18-sensor FOX 4000 Electronic Nose will be presented toillustrate both quantitative and qualitative applications.The ® nal results will then be presented on an opti-mized six-sensor GEMINI system. Several applicationsin the polymer and petrochemistry with the GEMINIElectronic Nose assess product integrity and provide astatistical quality/process control (SQC/SPC) chart inproduction facilities. The performances of the systemwill be addressed in terms of accuracy, reproducibilityand sample throughput.

Data acquisition and analysis software for gelpermeation chromatography with single or multi-detector capabilities

Stephen O’Donohue, Elizabeth Meehan, Geo¡ Cowell and GregSaunders, Polymer Laboratories Ltd, Essex Road, Church StrettonSY6 6AX, UK

In contrast to regular HPLC applications, the softwarerequirements for data analysis in gel permeation chro-matography (GPC) are very speci® c. In the simplestcase, using a single concentration detector, well-estab-lished algorithms can be applied. Additional detectorsadd complexity to the data analysis and any GPC-speci® c package must be ¯ exible enough to address avariety of multidetector applications. This presentationreviews the philosophy of an approach to addressingthese requirements in a new GPC software suite, Cir-rusTM. Mathematical functionality is obviously a pre-requisite and the capabilities of single and multidetectorsoftware modules will be illustrated. However, otherimportant features such as ease of use, database function-ality and traceability will also be discussed.

Fast-automated characterizatio n of polymersusing pyrolysis gas chromatography/mass spec-trometry

Christopher R. Mubarak1, Stephen L. Morgan1 and Russell W.Zeigler2, 1University of South Carolina, Chemistry and Biochem-istry, 631 Sumter Street, 1212, Columbia, SC 29208-0001, USA,


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2University of South Carolina, Chemistry, 631 Sumter St Csrc,Rm 212, Columbia, SC 29210, USA

Achieving fast analysis of polymers remains an increas-ingly important task for many routine laboratories.Applications range from assessing stability to basicstructural characterization. Analytical pyrolysis, inwhich gas chromatography (GC) or GC/mass spectro-metry (MS) is coupled to a pyrolysis reactor, is acommon approach for rapid polymer analysis. However,the GC separation itself is often the limiting step indecreasing overall analysis time. We have converted aHP-5890 Series II GCD quadrupole GC/MS into a high-speed separation instrument using a Thermo OrionEZ-Flash (TM). The column is resistively heated attemperature ramps as fast as 20 8C/s. Finally, by couplinga CDS Analytical Model 2500 Pyrolysis Autosampler tothe GCD inlet, up to 45 samples can be analysed in anautomated fashion. Analyses that conventionally re-quired 30 min or more are now completed in 2± 5 min,depending on the application. Because of heavy caseloadrequirements, forensic laboratories are continually look-ing for ways to increase sample throughput. As anexample of fast polymer characterization for forensicanalysis, we have investigated the possibility of automo-bile tire analysis. Rapid analysis of automobile tires couldprove to be very useful for law-enforcement agencies inmatching questioned evidence, such as tire residue, to asuspect tire. Even if such information cannot providede® nitive identi® cation, information from tire character-ization may serve to reduce the number of possibilities ina forensic investigation. The combined EZ-Flash (TM)/pyrolysis autosampler/GCD instrument greatly increasessample throughput while producing data of suæ cientquality to discriminate tires from diå erent manufac-turers.

Automated plasma protein precipitation and� ltration

Sha Liao1, Sarah Johnson1, Susan Dawson1 and Roger Roberts2,1Hamilton Company, Instrument Marketing, 4970 Energy Way,Reno, NV 89502-4178, USA, 2Ansys Technologies, Inc., 25200Commercentre Drive, Lake Forest, CA 92630-8810, USA

Biological sample preparation, particularly by removalof plasma protein, is an important step in drugmetabolism and pharmacokinetic studies. Currently,most sample preparations are done manually usingsolid-phase extraction in low-density formats, or usinga time-consuming centrifugation process. Automatingplasma protein precipitation will dramatically increasethe throughput of drug discovery. While some automatedworkstations have the capability of excellent liquidhandling with air-displacement pipetting, the cloggingtips caused by aspirating precipitated protein hamperstheir use in this application. Clogged wells have alsobeen a problem for ® ltration of particulate materialusing the traditional 0.45-mm ® lter plate. In thispresentation, a unique system combining positivedisplacement pipetting, disposable tips and vacuum,completely automates the process of preparing theparticulate free, ready for use plasma in a 96-well format.This system uses a MICROLABaª AT plus 2 automatedrobotic pipetting system and CaptivaTM 96-well depth

® lter plates with a CaptiVacTM Vacuum Collar (Ansys,Lake Forest, CA, USA) to overcome dripping andclogging problems. This study presents a throughputcomparison between the manual and automated pro-cesses, volume of ® ltration recovery and two standardsrecovery.

Generic techniques in chiral method development

Denise M. Wallworth and Thomas E. Beesley, AdvancedSeparation Technologies, Inc., 37 Leslie CT Box 297, Whip-pany, NJ 07981-1635, USA

The process of using short, coupled columns for fastgeneric screening in chiral method development isexamined. Rapid results in three diå erent types ofmobile phase systems are achieved enabling selectionof a method according to the need. Molecules with abroad range of diverse stereogenic environments candemonstrate selectivity in < 2 h. Mobile phase design iscritical to establishing the window of selectivity for verydiverse racemates. This presentation will focus on the keyissues involved in this design and present the rapidprotocols for optimization of the ® nal separation. Ex-amples of screening several diå erent molecular classeswill also be given.

Automated hit selection and consolidation

Linda Aubin, Mike Dameron, Paul Lefebvre and Ray West,Gilson, Inc., Center for Integrated Discovery, 701 GeorgeWashington Highway, Lincoln, RI 02865, USA

Much of the modern drug-discovery process is carriedout in groups or sets of compounds, from compoundsynthesis to biological screening. The group formatis often a microplate but may also be a set of tubesor vials. As compounds move through the processthey are eliminated from consideration or advancedas ` leads’ . The lead compounds are consolidated atvarious points in the process based on speci® edcriteria and then moved on to the next step in theprocess. This process has been referred to as ` cherry-picking’ or ` hit-picking’ and is used to focus in onthose compounds that are most likely to be viable drugcandidates. Automated liquid handlers that can be usedto consolidate compounds are important tools toimprove the productivity of the process. For example,consolidation of synthetic compounds that passquality control or positive hits from biological screeningare applications. Three automated hit-picking systemswill be compared here in terms of cost, throughput andcapacity. The comparison will include system usingdisposable tips and ® xed probes, single-probe andvariable span, multiple-probe liquid handlers. The deckof the liquid handler is de® ned based on the source anddestination plates or vials. Methodology is created tooptimize such features as transfer amount, throughputand probe rinsing. Samples for consolidation are chosengraphically from the source plate layout or by import-ing a list from the user database. Upon completion, theuser can generate a work list report.


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Automation of dynamic liquid-phase micro-ex-traction in determination of polycyclic aromatichydrocarbon

Li Hou1 and Hian Kee Lee2, 1Department of Chemistry, LowerKent Ridge Road, 3 Scien, Singapore 119260, 2Department ofChemistry & Biology, Provo, UT 84602, USA

An automation of dynamic Liquid-Phase Micro-Extrac-tion (LPME) technique combined with HPLC has beendeveloped for the extraction of polycyclic aromatichydrocarbon (PAHs) from water samples. In this study,the main objectives are: (1) automating the experimentwith a Harvard (PHD2000) apparatus so that theexperiment could be simpli® ed; (2) obtain a largeenrichment factor by increasing sampling volume andnumber of samplings; and (3) improve the precision.The factors in¯ uential to dynamic LPME were opti-mized with automation. The eå ects of organic solvent,plunger movement pattern, sampling volume, numberof samplings, salt concentration and temperature wereinvestigated. Analytically, the relative standard devi-ation is < 6%. Enrichment factors could reach over280. By using a sampling volume of 20 ml and 40samplings, the limits of detection were between 0.35and 0.60 mg/l.

Regulatory control and monitoring of contami-nants and residues in Nigeria

Momodu S. Momodu, Dora N. Akunyili and S. Agegbu Denloye,Federal Secretariat Phase 11, Lagos, PMB 12949, Marina,Nigeria

The control of persistent organic pollutants which areproblematic in many developing countries has receivedadequate attention in Nigeria with the use of TLC, GCand HPLC techniques in NAFDAC laboratories. Resi-due data of organochlorines , organophosphorou s andcarbamate pesticides have been obtained. Impressivereports on the quality of formulations and quantities ofresidues from studies carried out so far indicate thatcapacity building in the ® eld of contaminants andresidues control is worthwhile. The routine evaluationof toxic metals in food, water and environmental samplesusing atomic absorption spectroscopy has helped in thesetting of standards and national regulations. The scopeof monitoring and control is being extended to mycotox-ins; and veterinary drug residues with the prospect of useof ELISA kits. Data acquisition for regulatory control topromote consumer safety and international trade isexpected from these studies.

Long-term monitoring of nitrate in the Tangipa-hoa River

Jeremy S. Stevens1 and John R. Allen2, 1Southeastern LouisianaUniversity, Chemistry and Physics, SLU 878, Hammond, LA70402-0001, USA, 2Southeastern Louisiana University, 612Oakhall Drive, Holly Springs, NC 27540-8719, USA

The Tangipahoa River is an important river in theparish of Tangipahoa, Louisiana, USA. The river startsas far north as Liberty, Mississippi, and empties intoLake Ponchatrain. Unfortunately, the river has been

prohibited to swimming and ® shing due to environ-mental reasons. This research deals with the analysis ofnitrate, which plays a role in the nitrogen cycle. Thereare many dairy farms that run along the river andsewage from homes which can be an important sourceof nitrate, a fertilizer for plants. At high concentrationsnitrate is harmful to ® sh and man. Seven sampling sitesfrom Tangipahoa Parish were chosen with six sitesbeing highways that cross the river and the other by alocal boat launch. Each sampling site was 3± 5 milesfrom the next. Two representative samples were takenfrom each site along the river. The samples were thentaken back to the laboratory for analysis by an ion-selective electrode for nitrate. This will give an indica-tion of the ¯ uctuations of nitrate over time. These¯ uctuations are compared with weather patterns duringthe time of testing. Under normal weather conditions,test results show that the concentration of nitratedecreases as it approaches Lake Ponchatrain. After aheavy rain, the concentration rapidly increases in areasthat are normally low. This may be due to an increaseof nitrate from the runoå from local dairy farms orsewer drainage.

Characterization and evaluation of kaolin treatedwith ultrasonic energy and chemical agents in thehydrodesulfurizatio n of a vacuum gas oil

Alfredo Castillo-Mares, Rebeca Silva-Rodrigo, GuillermoSandoval-Robles, Aaron Melo-Banda and Ricardo Garc|è a-Ala-milla, Instituto Tecnoloè gico de Ciudad Madero, Ingenier|è aQu|è mica y Bioqu|è mica, Juventino Rosas y Jesus Urue, CdMadero, Tamaulipas 89440, Mexico

In the present work, kaolin was submitted to the action ofchemical agents, such as HCl, NH4Cl, HNO3 andNH4Cl2M. solutions and treated with ultrasonic energy(US) or magnetic stirring and used as support of NiMocatalysts. Aluminium extracted was measured by atomicabsorption while the acidity of the treated material wasdetermined by the dehydration of isopropyl alcohol andpotentiometric titration with n-butylamine. Surfaceproperties were determined with an N2 physi-sorptionand the structures by X-rays diå ractograms. Hydrode-sulfurization (HDS) was carried out in a batch reactor at350C and 80 kg/cm2 initial H2 pressure for 3 h. Sulfurremoved was measured with an CHNOS analyser.Activation with chemical agents caused a dealuminationbeing deeper when kaolin was acid treated than with thederived salt. The US eå ect reinforced this and deal-umination values were close to 11 wt%. Total acidityfrom n-butylamine varied from 1.70 to 1.06 meq/g corre-sponding the highest values to the samples treated withUS energy. Only propylene appeared in the chromato-grams from the isopropyl alcohol dehydration indicatingBronsted type sites. The highest surface area was for theHNO3 US treated sample being almost 10 higher thanthe raw material. Total pore volume varied from 0.051 to0.139 cm3/g and pore diameter from 208 to 46 A® . X-raysdiå ractograms showed a more amorphous material whenUS energy was used. US treated kaolin with HCl gave49% HDS compared with a 14 and 34% with HNO3 anda commercial catalyst, respectively.


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Quality control of a wheat gluten hydrolysate foruse as a raw material in cell culture media

Heather K. Schwartz1, Matt Caple1, Kevin Ray1, Ben Cutak1

and James Blasberg2, 1Sigma-Aldrich, 3300 S. 2nd Street BldgN3, Saint Louis, MO 63118-3306, USA, 2Sigma-Aldrich,Mallinckrodt & 2nd Streets, St Louis, MO 63147, USA

Animal cell culture medium is a mixture of amino acids,vitamins and salts which is traditionally supplementedwith serum or other animal-derived components. Proteinhydrolysate s prepared from a variety of sources includinganimal tissues, milk protein and plant tissue have beenshown to be capable of replacing or reducing the need formore expensive components such as serum. More re-cently, due to increasing concern about the potentialcontamination of animal-derived components by adven-titious agents, plant hydrolysate s have been used toreplace animal source medium components. In particu-lar, we have found a wheat gluten hydrolysate, optimizedfor high levels of small peptides and rich in stableglutamine, to have particular value as a component inthe formulation of serum-free cell culture medium.A decrease in productivity, with no corresponding de-crease in cell growth, has been associated with lot-to-lotvariability of this hydrolysate. We have developed anRP-HPLC method with photodiode array and MSdetection for peptide mapping of wheat gluten hydro-lysate in order to examine lot-to-lot variability. Signi® -cant diå erences noted in the pro® le are correlated toobserved diå erences in the biological activity of cellsgrown in medium containing diå erent lots of hydroly-sate. The presentation will demonstrate an LC-PDA-MSmethod that can be used as a quality control method forwheat gluten hydrolysates .

Search algorithms in an integrated analyticalsystem for searching databases of spectra andchemical structures with improved searching re-sults

Michael Boruta and Marie Scandone, Bio-Rad Laboratories,Informatics Division, Sadtler Division, 3316 Spring GardenStreet, Philadelphia, PA 19104-2552, USA

All IR search systems for spectral interpretation andstructure elucidation of organic compounds depend onthe same principles. In them, a spectrum of an unknowncompound is compared with all spectra in a referencedatabase and the best matches are reported. Mostsystems assign a degree of similarity, which re¯ ects thespectral properties common to the spectrum of theunknown compound and the reference compound. Forall the reference compounds showing a degree of simi-larity, a chemical structure can usually be listed. Thispresentation will examine how a classi® cation search canpoint to a class of compounds when an identi® cation isnot possible. Such a system can focus on the classi ® cationof the compounds listed and can control if an identity, asimilarity or a classi® cation search is performed toproduce the best results. With this approach, it is possibleto modify the search characteristics of a database searchsystem in a predictable manner and improve searchingresults. Thus, a user can tune the system for maximumperformance.

Process control of pharmaceutical products usingAOTF-NIR

Igor Nazarov, Brimrose Corporation, Spectrometer Division, 5024Campbell Blvd, Baltimore, MD 21236-5974, USA

Our discussion will be on the development of our systemto monitor and control processes of pharmaceutical prod-ucts in stages from raw material ID to ® nal productinspection. Our system uses various types of BrimroseAOTF-NIR Spectrometers, special operator ’ s interfacesand software. Our system can be used to monitorproduction of solid dosage forms, liquids and powdersin pharmaceutical and other industries. Advantages ofAOTF-NIR spectrometers will be shown and demon-strated. We will present the Brimrose family of the highlyreliable, fast (up to 16 000 wavelength s¡1) and on- andoå -line analysers including the FreeSpace Spectrometerfor non-contact diå use re¯ ectance measurements in` open’ § space or through windows, 16 channels multi-plexer for transmission, trans¯ ectance, diå use re¯ ectancemeasurements with diå erent kind of ® breoptics. The newproducts including miniaturized Luminar 3075-701,battery operated, spectrometer that rides on pharmaceu-tical blenders and controls the uniformity in real timeand the ` Universal’ Luminar 3070 Tablet Analyzer thatmeasure tablets on a moving belt in transmission anddiå use re¯ ectance modes will be demonstrated. TheDevelopment of new 21 CFR Part 11-complied softwareto back it up for the pharmaceutical industry will beshown.

Automatic HPLC method development: practicalexperiences with pharmaceutical and chemicalsamples

Wolf-Dieter Beinert1, Volker t Volker Eckert1, Reinhold Spatz1,Sergey Galushko2, Irina Shishkina3 and Vsevolod Tanchuck3,1Merck KGaA, Scienti¢c Laboratory Products, Frankfurter-strasse 250, Darmstadt D-64271, Germany, 2Institute of Bioor-ganic Chemistry National Academy of Sciences, Fine OrganicSynthese, Im Wiesengrund 49-b, Muehltal D-64367, Germany,3Institute of Bioorganic Chemistry of National Academy ofSciences, Murmanskaya 1, Kiev-94, Ukraine

ChromSword Auto is an automated expert system devel-oped to search for optimum conditions in reversed-phaseHPLC. Parameters that can be optimized are concentra-tion of an organic modi® er for isocratic or gradientconditions and temperature. For proper peak tracking,ChromSword Auto oå ers two possibilities. In the mostsimple mode, a standard of each compound to beseparated is required and the optimization procedureperformed with the single standards only. However, oftenfor all compounds to be separated suitable standards arenot available. This is usually the case with pharmaceu-tical quality-control samples for purity and stability tests.The advanced peak tracking mode of ChromSword Autoallows one to perform the main part of the optimizationprocess with the compound mixture itself and thenumber of standards necessary is reduced to only two,regardless of how many compounds are present in themixture. As result of the optimization procedure, thesamples are separated into a maximum number of peaks,with optimum resolution and minimum analysis time. By


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practical optimizations examples of real pharmaceuticaland chemical samples, the possibilities of this new auto-mated chromatography method development system areexplained.

Bioconnectors for discovery

Scott Deutsch and Malcolm McGregor, LabVantage Solutions,245 Highway 22, Bridgewater, NJ 08807-2560, USA

Discovery science presents a severe challenge to com-panies attempting to make use of emerging techniques.To maintain competitive advantage, it is imperative thatcompanies use cutting-edge technology as it becomesavailable. The result is a constantly changing environ-ment that is diæ cult for coordinating and manage-ment systems to keep up with. A typical year-longpurchase and implementation cycle that is common forlaboratory management software becomes untenableunder the circumstances found in discovery-focusedorganizations. Likewise, the in exibility of these systemsmakes them sluggish, diæ cult and expensive to change.The individual nature of each laboratory due to the largenumber of options available for each subsystem, i.e.instruments, robotics, expert software systems as well asthe diå ering requirements of the speci® c science, meansthat an out of the box solution is very diæ cult toimplement. In fact, commercial packaged software ven-dors have found it diæ cult to carve a niche in thismarket. This paper explores a novel modular approachto providing the required commercial software with thetools required to meet the strictures outlined above.Attention is paid to providing small pieces that can beadded and removed at will from the system. These aredescribed in the system as BioConnectorsTM. Advantagesinclude a rapidly implementable system that can beupdated continuously by laboratory personnel as re-quired. The paper also explores some of the challengesfaced in keeping the modules up to date and transferablefrom operation to operation as well as maintaining theability to support them commercially.

Construction of LIMS systems using analyticaldata system

Toshinobu Yanagisawa, Kiyoshi Yamashita, Yoshihiro Hayaka-wa, Atsushi Yoshida, Yasuhiro Funada and Teruhisa Ueda,Shimadzu, Analytical Instruments Division, 1 Nishinokyo-Ku-wabaracho Nak, Kyoto 604-8511, Japan

Construction of LIMS systems often requires developingcustom-made software. However, many automation fea-tures such as sample sequence, data export and OLEautomation have been implemented in an AnalyticalData System. For example, more than 1000 samplescan be analysed in a single HPLC system with anautosampler. Remote automation is easily achieved byusing an OLE automation feature with Visual BASIC orother programming languages. The user interface can becustomized for a speci® c purpose of application. One ofproblems in such a system is lack of integrity of data fromdiå erent instruments such as HPLC, GC, LCMS,GCMS, UV, FT-IR and electric balance. It is not rarein a laboratory to analyse a sample with multiple

instruments. Shimadzu LabSolutions Analytical DataSystems solved this issue by CLASS-Agent data manage-ment software using a back-end database on a networkserver. Oracle, MS-SQL server or MS-ACCESS can beused for the database. Raw data and calculation resultsfrom each instrument are stored to the database togetherwith sample information such as sample name andsample ID. An application programme can be launchedto perform custom calculations when each instrumentstores data to the database. As data with the same sampleinformation are easily queried, analysis results of thesample with multiple instruments can be browsed inthe same platform. These results can be summarized toa single report. A generic interface to store data to thedatabase is supported for multivendor instruments. Thesefeatures contribute to enhance throughput in a labora-tory. The focus of this presentation is to implement LIMSsystems eå ectively by using current analytical datasystems.

The bene� ts of automation with a LaboratoryInformation Management System (LIMS) imple-mented at a chemical facility

Mathew Abraham, Tom Miller and Danny Cormier, AcceleratedTechnology Laboratories, Inc., Engineering, 496 Holly GroveSchool Road, West End, NC 27376-8412, USA

Chemical testing laboratories are under increased pres-sures to track a variety of samples in a rapid yet thoroughway while complying with federal regulations andstandards. Coastal Chemical Co. serves the naturalgas processing markets as well as the re® ning, chemicaland petrochemical markets. The analytical laboratoryselected and implemented a Laboratory InformationManagement Systems (LIMS) to automate their login,enhance their data accessibility, incorporate a full chainof custody, implement a full audit trail, automaticallytrack their sample status, provide QA/QC functionality,automate reporting and manage the vast array oflaboratory data generated. This presentation will focuson the advantage s of tracking samples, viewing samplestatus real time, viewing charted data, statistics withSample Master Pro LIMS. The company required adata management system with great ¯ exibility to tracka myriad of samples and comply with federal regulationswith integrated reporting. This presentation will focus onthe LIMS selection criteria, implementation, and customreport creation. In addition, automation bene® ts realizedfollowing implementation will also be discussed.

Laboratory Information Management System(LIMS) that spans the network and beyond

Don Kolva, Tom Miller and Christine Paszko, AcceleratedTechnology Laboratories, Inc., 496 Holly Grove School Road,West End, NC 27376-8412, USA

In the past few years, a laboratory that was implement-ing an LIMS had to decide whether it wanted atraditional client/server con® guration or a browser-basedLIMS. The client/server con® guration is easier to modifyand provides a better user interface, but the browser-based system allows for intra- and Internet access to


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users without the need to install any client side software.For many laboratories, the ideal solution is to implementone system that uses both interfaces. With a singleLIMS that can use either con® guration, the laboratorycan choose the interface that is appropriate for theuser. For example, an internal user can use a client/servercon® guration to take advantage of the speed, reliabilityand ¯ exibility of an internal network. This con® gurationallows for better report modi® cations, data transfersto other applications such as Microsoft Oæ ce and amore traditional GUI interface (graphical user interface)for the user. Extending the same LIMS to an intra-or Internet allows remote users to view the informationglobally. It also provides a better interface formobile users who may only be able to connect to thesystem via the Internet. A browser interface also usessoftware that is probably already on the computer,making maintenance easier than the client/server con® g-uration. This presentation will explore the advantagesand disadvantages of each con® guration using real-lifeexamples and will show that you can have your cake andeat it too!

The role of users in an LIMS (Laboratory Informa-tion Management System) implementation

Lisa Goren£o, Jodie Roybal and Kim Paszko, AcceleratedTechnology Laboratories, Inc., 496 Holly Grove School Road,West End, NC 27376-8412, USA

The human element is most often ignored during anLIMS project, but some LIMS project failures (or lack offull implementation) are due to human issues and lack ofinvolvement by the users rather than technology issues.This presentation will look at ways of improving userinvolvement and hence increasing the probability ofsuccess for your laboratory ’ s LIMS project. This pre-sentation will help to answer the questions: How do youmake an LIMS project successful and what are thereasons for failure? How do you involve users activelyin your LIMS project? What are the styles of implement-ing your project involvement plan? How should youevaluate the types of users and match their involvementto phases of the LIMS project for best eå ect? Oftendiå erent departments within a laboratory face uniquechallenges that other groups do not encounter. It iscritical that the data management requirements of thesegroups are taken into account and with today’ s ¯ exibleLIMS system, special functions can be added and user’ sneeds can be accommodated. Often times the Informa-tion Technology group has special requirements to matchcurrent IT infrastructure, in-house expertise, securityrequirements and future growth plans. It is critical thatthe various groups within the laboratory cooperate andparticipate in the selection process through the imple-mentation. Those typically involved in the project teaminclude; the laboratory manager, the IT manager, thebusiness manager, QC group leader and there may beother departments. It is also helpful if one person canserve as the mediator, this person should understand theterminology and needs of all groups. A blueprint for asuccessful LIMS implementation with a focus on thehuman element will be described.

ONQ, a system for automated quality assurance(QA) of on-line NIR analysers

Kari Aaljoki, Automation Engineering, PO Box 310, PorvooFIN-06101, Finland

The success of on-line NIR-application depends onseveral factors that can be divided into three categories:sample handling (e.g. water and particle removal andtemperature stability); analyser itself (e.g. repeatabilityand accuracy of spectra, and S/N); and modelling (e.g.coverage of calibration sample space, quality of inputdata, and skill). When the number of analysers andmeasurements increases, it becomes obvious that somesort of automated system is absolutely necessary forroutine QA and continuous validation of the analyserfarm, because doing these tasks manually would simplybe too expensive. Without proper QA and validationNIR installations are bound to fail. The idea behindOnQ is simple. Spectra themselves contain a lot ofinformation about above-mentioned categories. OnQdoes rule based diagnosis of the spectra and SPC topinpoint hardware or sample handling problems. It alsouses SPC (Statistical Process Control) techniques to assessmodel performances based, e.g. prediction diå erences(lab-on-line result) and some multivariate measures ofprediction statistics and the validity of the model used. Itcollects also data (spectra and reference values) at thesame time for later model updates just to save time. OnQfeatures automated reporting, trending, and chartingfunctions for major key variables for easy visual inspec-tion. Problems are reported to maintenance peoplethrough e-mail. Technically speaking, OnQ has beenbuilt using Borland’ s Delphi 5 and it runs on WindowsNT. OnQ uses MS SQL server as its database engine.Laboratory data from in-house-built Oracle 7.3-basedLIMS running on HP9000 is collected using SQLqueries. Some reference values from other on-line analy-sers are collected from ABB’ s PMS (Process ManagementSystem) using its ODBC capabilities. This paper showssome simple examples taken from real situations in orderto demonstrate the usefulness automated QA and valida-tion of NIR-analysers.

On-line capillary electrophoresis instrument forindustrial processes

Stella Rovio1, Teemu Tyoë ppoë nen1, Pertti Vastamaë ki1, AriHokkanen1, Heli Sireè n

1 and Pekka Savolahti2, 1TechnicalResearch Centre of Finland, VTT, VTT Chemical Technology,Biologinkuja 7, Espoo FIN-02044, Finland, 2Technical ResearchCentre of Finland, VTT, PO Box 1401, Espoo FIN-02044,Finland

Capillary electrophoresis instrument for on-line controlof industrial processes has been developed withcontinuous ¯ ow delivery systems for solvent and sample.The instrument consists of a three partial cell unit, thecentre part for the capillary being changeable for¯ uorescence or UV detections and the front and rearmodules with channels for solvent control. The on-lineunit has ® xed tunnels for dynamic replenishment ofelectrolyte solution at its both ends. Owing to theon-line solvent delivery system constructed into the cellunit, the capillary and the platinum electrodes will be


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kept uncontaminated. Samples are introduced into thecapillary with pressure or electrokinetically or by using acombination of both the techniques. Samples with highionic strength will be on-line diluted with an electrolytesolution before they are introduced into high-speed CEanalyses. Voltages up to 2.5 kV can be used in separa-tions. This presentation demonstrates the work done onthe instrumentation and the advantages of dynamicsample and electrolyte feeding. Eå ect of ¯ ow conditionson pressure diå erences between capillary ends werestudied by determining the electro-osmotic ¯ ows withdiå erent electrolyte feeding rates and by comparingmigration times and peak shapes of separated samplepeaks from the electropherogram. Electrophoretic se-parations with laser induced ¯ uorescence detection(LIF) are demonstrated by using CAPS as backgroundelectrolyte for the analytes ¯ uorescein, mandelic acid andphenyl acetic acid. Preliminary results for the samplesoxalate, acetate and sulphate anion, detected by the UVdetector at 254 nm wavelength were also performed byusing OFM-PDC as a background electrolyte. On-lineapplications on pulp and paper process are demon-strated.

Computer system validation and electronic re-cords/signatures: a pharmaceutical laboratoryperspective

Sunday A. Brooks, Phyllis Hodson-Hutsell, Nancy Craig, TerryTripp, Bryan K. Jones and Sandy Birge, Eli Lilly & Co.,Tippecanoe Quality Control Labor, 1650 Lilly Road, Lafayette,IN 47909-9201, USA

What does computer system validation (CSV) andelectronic records/electronic signatures (ER/ES) meanto an analytical laboratory and its instrumentation?How is this process diå erent from laboratory GMPquali® cation of an instrument? In response to 21 CFRPart 11 (Code of Federal Regulations), manylaboratories are evaluating their laboratory instrumenta-tion for compliance. This federal regulation alsoassumes that validation exists for computer systems.Until the passage of Part 11, many laboratoryinstruments were not validated as computer systems. Thispresentation will detail the planning and execution ofthe CSV plan for Eli Lilly Tippecanoe Quality ControlLaboratory (QCL). Tippecanoe QCL and IT, guidedby a Quality Control CSV Coordinator, have completedthe analytical instruments’ computer system validations.Laboratory CSV requires signi® cant contributions fromchemists or other technical experts familiar with eachinstrument. Laboratory personnel must de® ne theinstrument requirements and execute a testing planwhich proves the capability of the instrument and itscomputer. Computer system validation of a laboratoryinstrument includes not only hardware/software reviewand testing, but also user training, system support andcontingency planning. Action and implementationplans for ER/ES will also be discussed. Plans can includesoftware upgrades, procedural controls, and physicalsecurity. Example plans for data integrity, securityand archiving for speci® c types of instruments will bediscussed.

Quanti� cation applications with sensor arraysystems: a new approach in quality control

Quitterie Lucas1, Vincent Schmitt2 and Tsung Tan2, 1AlphaM.O.S., Application Laboratory, 3 Avenue Didier Daurat,Toulouse F-31400, France, 2Alpha M.O.S., 20 Avenue DidierDaurat, Toulouse F-31400, France

The overall fragrance, ¯ avour and aroma intensityperception is very important for the consumer.Achieving complete customer satisfaction and royaltyrequires that the perceived odour to be excellent qualityand quantity all the time. These characteristics thenensure that luxury and premium brands maintain highpro® tability and market shares. The quality-controlaspects ensure that the once the fragrance or aroma hasbeen developed and demonstrated to meet the qualityrequirements of the consumer; in this presentation, wewill focus on one case study of quanti® cation: peppermintaroma in toothpaste. This industrial application willdemonstrate the ability of the Sensor Array System forthe quanti® cation of the level of peppermint and it alsodemonstrates how such information can be used easily inthe production environment to ensure constant qualityand quantity while ensuring minimum use of expensiveraw materials to achieve maximum results. The resultsobtained using calibration curve (PLS) and statisticalQuality Control (SQC) data for the quanti® cation andcontrol of peppermint quantity in toothpaste demon-strated the ability of FOX4000 and GEMINI SensorArray Systems for use in many industries where a highlevel of QC/QA is required to ensure satis® ed and loyalcustomers.

Automation of dilutions for various sample types:liquids, solids and di³ cult samples

Lynn Jordan, Jayne Brown and Stephanie Wilson, ZymarkCorporation, Zymark Ctr, 68 Elm Street, Hopkinton, MA01748-1668, USA

One of the most common operations in any laboratory isdilutions. Two common uses of dilutions are to preparestandards for calibration of analytical equipment, and toput samples into solution. In many laboratories,preparing standards and samples is a daily operation.Dilution of standards or samples is most commonly donemanually using volumetric ¯ asks. This method ofperforming a dilution yields an accurate result, butinvolves a manual operation, oå ers no documentationunless it is performed on a balance, and in manyinstances generates more sample or standard than isnecessary for the application. This poster will explorethe use of an automated workstation to perform dilutionsfor multiple types of samples, and provide 21 CFR Part11 documentation. Examples will be shown for dilutionsof solid samples, liquid samples as well as ` diæ cultsamples’ such as suspensions or creams. The manualmethod will be compared with an automated method,and the following parameters will be investigated: op-erator time, overall time to prepare samples and wastegenerated documentation of the dilution.


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Possibilities for automation of sample prepara-tion steps before LC or GC analysis using acommon autosampler

Eike Kleine-Benne1, Friedhelm Rogies1, Peter Popp2 and AndreaBuhr3, 1Gerstel GmbH & Co. KG, R&D, Aktienstrasse 232º232, Muelheim An Der Ruhr D-45473, Germany, 2UFZöUmweltforschungszentrum Leipzig-Halle GmbH, Sektion Analy-tik, Permoserstra E 15, Leipzig D-04301, Germany, 3FraunhoferInstitut fuë r Holzforschung, Wilhelm-Klauditz-Institut, Bienro-der Weg 54e, Braunschweig D-38108, Germany

Primarily autosamplers for chromatographic devices areused to increase the degree of utilization. A secondimportant reason to use autosamplers is the improvedreproducibility and comparability of analytical resultsbecause of their independence from diå erent users. Thisis accepted for sample injection and is important forsample preparation. A source of possible errors for samplepreparation steps is the operator. A software solution ispresented how a common autosampler for LC or GC(MPS, GERSTEL GmbH & Co. KG, Germany) can beused to avoid such errors without purchasing expensivelaboratory robots. A simple example is presented how toprepare standard solutions for daily calibration. A morecomplicated example shows a complete sample derivati-zation (esteri ® cation) followed by solvent extraction(toluene) and transfer of the extract in autosampler vials.A further application shows how the system in combina-tion with a membrane extraction inlet for headspace vialsis used for liquid± liquid extraction of chlorinated pesti-cides in aqueous samples with hexane. After extractionthe same autosampler is used to inject volumes of up to100 ml of the extract for GC analysis without further cleanup. In combination with a ¯ ow through cuvette and aLC injection valve an on-line coupling of a LC and a GCsystem is possible whereas the autosampler is used forinjection into the LC, fraction collection from the cuvetteand injection into the GC. A special application is thefully automated extraction of the GERSTEL Twisterwith acetonitrile± water mixtures followed by injectioninto an LC for analysis of PAH in aqueous samples.

Comparison of automated headspace techniquesstatic headspace, multiple headspace extraction,dilution and purge-and-trap using gas chromato-graphy

Ingo Christ, LEAP Technologies, PO Box 969, Carrboro, NC27510-0969, USA

Static headspace methods are widely used for volatile andsemivolatile compounds. They are environmentallyfriendly because they involve a solventless procedure.Many diå erent techniques have been developed to elim-inate the matrix eå ects of headspace methods, to simplifythem or to increase their sensitivity, reproducibility andthe speed of their analyses. Examples of these techniquesinclude Purge-and-Trap, Multiple Headspace Extraction(MHE) and a simple dilution series designed to create acalibration curve. Autosamplers can automate most ofthe steps required to perform these techniques; however,the fact that autosamplers typically are limited toperforming one speci® c technique is problematic in thatit is diæ cult to know which technique might work best for

each sample. Additionally, it is not easy to compare theresults of the analyses due to the diæ culty of transformingfrom one technique to another. This diæ culty is pri-marily caused by diå erences in sample handling andphysical variances in sample containers. This article willcompare the results of each technique and discusspractical applications. Since a uni® ed sample containerwill be used, comparison among the diå erent techniqueswill be possible. Using a standard sample in a standardheadspace vial, the results of a static headspace analysiswill be compared with a Purge-and-Trap method andwith MHE and sample dilution. All four methods will besoftware-controlled and performed without changinghardware. Sample preparation will be automated toeliminate manual error. Linearity and reproducibilityof the automated methods will be shown to comparefavourably with manual ones.

An automated approach to the removal of plasmaprotein precipitates by � ltration

Hung Nguyen, William Hudson, Roger Roberts and Dennis D.Blevins, Ansys Technologies, Inc., 25200 Commercentre Drive,Lake Forest, CA 92630-8810, USA

A new 96-well ® ltration product is evaluated on twoautomated liquid-handling platforms for the purposeof performing Plasma Protein Precipitation samplepreparation. The Captiva (TM) 96-well ® lter plateis an all-polypropylene depth ® lter with a minimumpore size of 0.45 mm that is quality controlled forcleanliness of materials and non-speci ® c binding of ana-lytes during ® ltration of plasma protein precipitates.The precipitating reagent and plasma samples weremixed and introduced into the Captiva 96-well ® lterplate by the automated platforms. The fast-¯ owing® lter design prevents clogging. The precipitate-freesamples were collected and submitted for analysis byHPLC. In this paper, two automated protocols forplasma protein precipitation ® ltration and supportingHPLC analytical data for the recovery of tricyclic anti-depressants and a comparison to typical plasma proteinprecipitation by centrifugation are presented. The datademonstrates a clean, fast, and eå ective automatedmethod of performing plasma protein precipitation® ltration.

Development of a multipurpose device (Auto-Shotsampler) for automated introduction of samplesinto the microfurnace type pyrolysers dedicated topyrolysis-gas chromatography (PY-GC)

Chuichi Watanabe1, Yoshio Kawahara1, Kunitaka Sato2, PaulTobias3, Hajime Ohtani4 and Shin Tsuge4, 1Frontier Labora-tories Ltd, R&D, 1-8-14 Saikon, Koriyama, Fukushima 963-8862, Japan, 2Frontier Laboratories Ltd, R&D, 61-2 Otsubo,Koriyama, Fukushima 963-0201, Japan, 3Quantum Analytics,Inc., 363 Vintage Park Drive, Foster City, CA 94404-1135,USA, 4Nagoya University, Department of Applied Chemistry,Chikusa-Ku, Furo-Cho, Nagoya, Aichi 464-8603, Japan

Automated sample introduction for PY-GC is playing avery important role in the analysis operations especiallyin the quality or process control of polymeric materials.


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The automation of the measuring system greatly en-hances the eæ ciency of analysis by saving labour andminimizes analytical errors caused by human manipula-tion. In the last Pittsburgh Conference, we have pre-sented a device, Auto-Shot Sampler AS-1020E, directlyattached to a microfurnace type pyrolyser of PY-2020iD(Frontier Lab, Japan). This automated sample intro-duction device was only applicable to the ¯ ash pyrolysisof up to 48 samples at a given pyrolysis temperature.This paper reports another capability added to theprevious features since then. The new feature is thatthe sample cup can be moved up and down verticallybetween the centre of the microfurnace of the pyrolyserand the waiting position in the Auto-Shot sampler bythe push of the pressurized carrier gas. With thiscapability, two analytical functions, stepwise analysis(combination of thermal desorption and pyrolysis),and heart-cutting GC analysis (correcting and analysingany desired portion of evolved gas from the sampleduring temperature programming) became eå ective.This entire operation can be automatically controlledindependently for each of the 48 samples mounted in asample tray by specially con® gured software installed ina personal computer.

An automated 96-well one-does-all extractionmethod for drugs of abuse in oral � uid

William C. Hudson1, Dennis D. Blevins1 and David O. Hall2,1Ansys Technologies, Inc., 25200 Commercentre Drive, LakeForest, CA 92630-8810, USA, 2Ansys Technologies, Inc., 605Vernon Tharp Street, Columbus, OH 43210-4007, USA

Automating sample preparation provides an eæ cientmeans to free staå from mundane tasks, which areprone to processing errors. With modern roboticsystems, high-throughput laboratories can dedicate ex-pensive equipment to perform a single analysis in arepetitive fashion. The key advantages include samplethroughput and traceability. With large roboticsystems, personnel are dedicated to software develop-ment and systems integration, providing the support tomake the automation requirements cost eå ective. Thecombination of a ` one does all’ extraction on anautomated platform facilitates the practicality of asmall laboratory investing in an automated system. Inthe past, diå erent drug types have required diå erentextraction protocols. On an automated system, eachdrug type would require a unique extraction pro-gramme and set of solvents. Combining diå erent drugtypes into one extraction protocol allows a laboratoryto simplify all extraction protocols (saving time) andstandardize the use of solvents and reagents (savingmoney). The con® rmation of drugs of abuse in oral¯ uid presents several challenges to clinical/toxicologicallaboratory analysis. The lack of available sample,compared with urine, and meeting the cut-oå levelsdrafted by SAMSHA, increase the diæ culty inperforming the con® rmation of multiple drugs in onesample. By developing an extraction protocol of12 major drugs of abuse (amphetamine, metham-phetamine, MDA, MDMA, MDEA, PCP, cocaine,benzoylecgonine, THC, codeine, morphine, and6-monoacetylmorpine) , it becomes possible to report

multiple drugs from one analysis and simplify multiplelaboratory protocols into one analysis. The sample wasextracted using an ANSYS Technologies SPEC-Ora-Lab-96-Well plate in a Hamilton AT 2 Plus roboticsystem. The Hamilton robot performs all sample pre-treatments, transfers and washes on an automatedplatform. The sample and wash volumes used wereadapted to the Hamilton’ s low volume pipettes tominimize sample aspiration and dispensing. Sampleswere subsequently concentrated, derivatized byMSTFA and analysed by GCMS. Samples were eval-uated by extraction recoveries (most absolute recoverieswere approximately 80%), and precision and accuracystudies were run over 3 days period. This presentationwill summarize the automated protocol and describelaboratory timesavings versus manual extraction proto-cols.

Using an automated vacuum system for 96-well� ltration and extraction

Susan C. Dawson, Sarah Johnson and Sha Liao, Hamilton Co.,4970 Energy Way, Reno, NV 89502-4178, USA

The key to high-throughpu t solid-phase extraction (SPE)and ® ltration applications is automation of the 96-wellmicroplate format. The Hamilton Co. MICROLABAutomated Vacuum System integrates a motorizedvacuum manifold and a vacuum controller into anautomated liquid-handling workstation. A variety ofcommonly available 96-well ® lter plates and collectionplates are used to prepare biological and other samplesvia SPE or ® ltration. The vacuum box is motorized sothat the ® lter plate can be transported from a wastechamber to an elution chamber. The manifold is com-pact, and two can be used in parallel on the deck of thepipetting workstation. Filter plates can be placed onto orremoved from the system manually or with a roboticarm. The vacuum controller uses feedback monitoring tomaintain the vacuum at the user-de® ned set point. In thispresentation, applications and analytical results of ® ltra-tion and extraction using this vacuum system will bedescribed.

Automated acid re� ux cleaning method for Te� on,glass and quartz accessories used in trace analysis

Robert Richter, Milestone, Inc., Department of Chemistry andBiochemistry, 160B Shelton Road, Monroe, CT 06468-2545,USA

Analytical chemists take numerous precautions toensure the lowest possible analytical blank. Theseprecautions include the use of high-purity acids andreagents for sample preparation, powder-free gloves,lint-free protective clothing, rigorous cleaning of allsample preparation and instrument components, etc.Despite all of these precautions, many chemists stillhave problems controlling the analytical blank. Theirproblems usually results from the way the samplepreparation and instrument components (i.e. Te onbottles, volumetric ¯ asks, glass and quartz ICP/ICP-MS spray chambers and torches, sample and solutioncontainers, microwave vessels, etc.) are cleaned and


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stored. Milestone has developed the TraceCLEANsystem to address these problems. The TraceCLEANsystem is unique in that is uses sub-boiling distillation toclean various sample preparation and instrumentcomponents. Sub-boiling distillation is the sametechnique used to prepare high-purity acids used foranalysis. This means components are not contaminatedby the acid used to clean them. Another importantfeature of the TraceCLEAN system is that the cleaningand drying of the components occurs in a sealedcontainer. This minimizes the amount of airbornecontamination that can be deposited on the compon-ents, keeping them clean until they are needed. Thisposter compares the cleaning of TFM microwave diges-tion vessels and high-purity quartz inserts using theTraceCLEAN, with the traditional method of cleaningthese components.

Interstage monitoring: the only way to guaranteethe purity of ultra-pure water

Paul Whitehead and Alan Mortimer, ELGA, R&D, HighStreet, Lane End, High Wycombe HP14 3JH, UK

All ultra-pure water systems include a de-ionizationpuri® cation step based on ion-exchange resin. Thisposter will describe the major bene® ts to be gained bysplitting the de-ionization system into two stages andmonitoring the water purity interstage. Interstage resis-tivity monitoring enables the ® rst ion exchange cartridgeto be replaced when its exchange capacity is exhaustedwhile the second stage is virtually unused. This ensuresthat all water passes through fully regenerated ionexchange resin before dispense, guaranteeing the waterpurity. This approach eliminates dependence on accurateproduct water resistivity monitoring, which is known tobe diæ cult and costly, and the reliance on frequentchecks on the outlet water quality. Ion-exchange resinutilization is maximized, the cost of cartridge exchange isreduced and the release into the product water of weaklybound components, such as organic compounds, silicaand boron, are avoided. Dividing the de-ionization intotwo stages also enables the optimum location of theultraviolet chamber interstage and permits the use ofcost eå ective integral TOC monitoring.

Speciation of mercury using atomic � uorescencespectrometry

Derek W. Bryce, Warren T. Corns and Peter B. Stockwell, PSAnalytical Ltd, Arthur House Cray¢elds Industrial Estate, MainRoad, Orpington BR5 3HP, UK

It is common knowledge that the toxicity of mercury isextremely dependant on the form in which it is present.In particular, organic mercury species are for moretoxic than the inorganic forms. There is a great needto be able to fully speciate mercury in order to gainfurther knowledge of its toxicity transformation,transportation and bioavailability. The two mostfrequently taken approaches to mercury speciationinvolve either HPLC or GC separation followed byelement speci® c atomic spectrometric detection.

Atomic ¯ uorescence spectrometry is an ideal detectorfor use in such systems as it shows unrivalled sensitivity,linearity, stability and freedom from interferences. Itis also extremely easy to couple to either HPLC or GCsystems, and is economical to run and maintain. Thepaper will describe two approaches for the speciationof mercury using atomic ¯ uorescence detection. The® rst system, using reverse phase HPLC-atomic¯ uorescence spectrometry allows the speciation ofmercury in aqueous samples without the need for anyderivitization or extraction into organic solvent.On elution from the column, UV oxidation is usedto convert organic mercury to Hg2 + before reductionto Hg0 with SnCl2. A preconcentration step may alsobe employed to reach lower levels. The secondapproach uses GC separation of mercury species usinga non-polar megabore capillary DB-1 column. As thespecies elute from the column, they pass through apyrolysis unit to convert the mercury to Hg0 before itenters the AFS detector. Advantages and disadvantagesof each system will be reviewed and extractionprocedures for a range of sample types, presented, alongwith results for a range of samples and certi® edreference materials.

Atomic � uorescence: 15 years’ experience of thetechnique as a measurement tool for mercury inlaboratory and process stream applications

Peter B. Stockwell, Warren T. Corns, Derek W. Bryce andJasmina Allen, PS Analytical Ltd, Arthur House Cray¢eldsIndustrial Estate, Main Road, Orpington BR5 3HP, UK

Over the past 15 years there has been considerableregulatory activity relating to the determination ofmercury in both laboratory and process applications.PS Analytical and its academic and industrial partnershave been actively involved in developing the chemicalprocedures required to measure accurately and reliablythe levels of mercury in many matrices. Procedures areavailable for all European and US (EPA) regulatoryneeds for including EPA 1631, 245.7 and CEN PrEN13506. Procedures involving continuous ¯ ow analysis,discrete sample analysis and amalgamation coupled tothese procedures will be described with particular refer-ence to the matrices for which each is particularly useful.The sensitivity, linearity and ruggedness of the techniquealso makes it an ideal approach for process monitoringapplications. However, these various applications pro-vide extremely diæ cult analytical applications. Severalapproaches to solving particular issues relating to incin-erator waste, chlor-alkali and sulfuric acid matrices willbe described in detail.

On-line continuous � ow liquid membrane extrac-tion-C18 precolumn-based trace enrichment forthe determination of sulfonylurea herbicides inwater by liquid chromatography

Gui-Bin Jiang1,2, Jing-Fu Liu1 and Jing-Bo Chao1, 1ResearchCentre for Eco-Environment, PO Box 2871, Beijing 100085,P. R. China, 2Faculdade de Ciencias Farmaceutc, BragancaPaulista 12900-000, Brazil


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Coupling on-line a continuous ¯ ow liquid membraneextraction (CFLME) device and a short C18 precolumnwith HPLC, a novel automatic system was developed forthe trace determination of sulfonylurea herbicides inwater. After preconcentration by CFLME, which is thecombination of continuous ¯ ow liquid± liquid extractionand support liquid membrane (SLM) extraction, thetarget analytes were enriched in 950 ml of 0.5 MNa2CO3-NaHCO3 (pH 10.0) buå er used as acceptor.This acceptor was neutralized and transported to a C18precolumn (30 £ 4.6 mm) where analytes were absorbedand focused. Then the focused analytes were injectedonto a C18 analytical column for separation and detectedat 240 nm with a diode array detector. Metsulfuronmethyl (MSM), sulfometuron methyl (SMM), andDPX-A 7881 were separated with a mobile phase consistsof methanol and acetic acid± sodium acetic buå er at a¯ ow rate of 1 ml min¡1. With an enrichment time of40 min and enrichment sample volume of 80 ml, theenrichment factors for these three sulfonylurea herbicidesare about 1000. This proposed method was applied todetermine sulfonylurea herbicides in river water, sea-water, lake water, tap water, bottled mineral water.Compared with the existing SLM-short C18 precol-umn-HPLC method, this proposed procedure has theadvantages of higher system stability, higher enrichmentfactor with shorter analytical time, and lower detectionlimits.

Automation of multidimensional chromatographyfor protein puri� cation in high-throughput struc-tural studies

Thomas Pless, Paë r Eklund and Jill A. Sigrell, AmershamPharmacia Biotech, R&D, Bjoë rkgatan 30, Uppsala SE-75184, Sweden

Structural genomics, also known as structuralproteomics, has taken oå . To determine structures,by both X-ray crystallography and NMR, fairly largeamounts (> 10 mg) of target protein in a pure andhomogenous form are usually required. Therefore,simple high-throughput puri® cation of proteins is neededby structural biologists. The ¯ exible A« KTA platformcan be con® gured in diå erent ways to suit preferredpuri® cation schemes. By using a new con® guration onA« KTAexplorer, supplemented with two seven-portvalves and two sample loops, automated two-steppuri® cations of diå erent proteins can be performedserially. Soluble, well expressed, proteins taggedwith either (His)6 or glutathione S-transferase (GST)were used in the evaluation of this new con® guration.Two diå erent puri® cation schemes were compared: (1)aæ nity chromatography followed by gel ® ltrationand (2) aæ nity chromatography followed by anintermediate desalting step and ion exchangechromatography. Both methods have the capacityto produce up to 50 mg of 90± 99% pure targetprotein. With this new con® guration, the researchersaves time and reduces protein losses comparedwith the same chromatography steps performedmanually.

Large-scale protein identi� cation using automatedgel processing and high-throughput MALDI massspectrometry

James Langridge1, Je¡ Brown1, Ronan O’Malley1, AlistairWallace1, Dominic Gostick1 and Philip Young2, 1MicromassUK Ltd, Life Sciences, Floats Road, Manchester M23 9LZ,UK, 2Micromass UK Ltd, Department of Chemistry, PO Box904, Emory, VA 24327-0904, USA

The huge increase in genomic sequence informationavailable has allowed the large scale, high-throughpu tidenti® cation of proteins (usually separated by 2DPAGE) by mass spectrometry, Matrix assisted laserdesorption ionization (MALDI) time of ¯ ight massspectrometry provides an ideal method for protein iden-ti® cation usually with fmole levels of sample where theacquisition and processing of the data can be easilyautomated. The provision of samples for high-through -put automated MALDI TOF mass spectrometry hasbeen addressed by the introduction of robotics. By usinga gel-cutting robot, excision of single or multiple 1 mmdiameter gel cores into a 96-well microtitre plate well canbe easily achieved. The software image of the gel is thenannotated with the excised spot number and tracks thesample position in the microtitre plate. The microtitreplate can be transferred to an automated in-gel digestionrobot which performs reduction, alkylation, in-gel trypticdigestion and peptide extraction for the gel samplesfollowed by automated spotting of the peptides onto aMALDI target plate. This robot is totally enclosed toprevent external contamination of the gel samples. In thispaper we describe a bench-top MALDI mass spectro-meter ® tted with an automatic plate-changing robot. Alldata acquisition, processing and searching is performedin an automated mode (unattended operation). Theresulting protein identi® cations can then be directlylinked with the gel image. This approach will be dis-cussed, with speci® c examples obtained from the largescale analysis of samples originating from 2D gel electro-phoresis.

DNA hybridization detection at ultratrace level by� ow injection surface plasmon resonance

Fayi Song, Jianqiao Lin and Feimeng Zhou, California StateUniversity, Chemistry and Biochemistry, 5151 State UniversityDrive, Los Angeles, CA 90032-4226, USA

A ¯ ow-injection device is combined, through the use of alow-volume thin-layer ¯ ow cell, with a sensitive surfaceplasmon resonance (SPR) spectrometer equipped with abiocell photodiode detector. The preliminary resultsshowed that the resultant ¯ ow injection SPR (FI-SPR)device could be applied for high-throughput, sequence-speci® c DNA analyses at ultracetrace levels. Self-as-sembled monolayers (SAMs) of a 30-base oligonucleotidewhose 30 end is derivatized with a mercaptohexyl tethergroup (HS-ss-ODN) were immobilized onto the thin Au® lm and then used as a model heterogeneous sensor forthe analysis of a 47mer target. The DNA hybridizationassay based on this FI-SPR device was also found topossess a high sample throughput and excellent samplespeci® city. The sensitivity of the FI-SPR will be com-


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pared with that of other types of SPR and severalultrasensitive techniques for labelled DNA targets (i.e.¯ uorescence-tagged and radiolabelled DNA samples).The use of a hexylamine-terminated 30mer probe cross-linked onto a preformed SAM of thiolated succinimi-dylundecanoate will be tried to further decrease thedetection level. The possibility of using SPR for detectionof a small number of ODN molecules will be exploited.An application of detecting Arabidopsis thaliana mRNAexpression pattern during transcription process will beattempted using this system.

Development of continuous monitoring system forgas generation behaviour during coking reaction

Masayuki Nishifuji, Koji Saito and Yuji Fujioka, Nippon SteelCorporation, Advanced Technology Research Lab, 20-1 Shintomi,Futtsu, Chiba 293-8511, Japan

To characterize coking reaction of coal in iron-makingprocess, it can be important information to realize gasgeneration behaviour on pyrolysis of coal. In this study, anew monitoring system for gas generation behaviourduring coking reaction has been established. This systemconsists of three parts, a carrier and atmosphere gassupply line, a reaction tube (heating a sample with anelectric furnace) and two detectors of Fourier-transforminfrared spectrophotometer (FT-IR) and of a sensorfor hydrogen which cannot be detected by FT-IR.Consequently, this system enables one to monitor allgenerated gases simultaneously. Coal powder sample innitrogen gas ¯ ow was heated, and generated gases weredirectly lead to detectors with nitrogen gas carrier. Thismethod has a good time-resolution of roughly 10 s withan improved cell without staying of gas ¯ ow, so that ashort time reaction of coal pyrolysis can be monitored.Using this system, diå erent gas pro® les of two typicalcoals during coking reaction, which have quite diå erentcharacters in iron-making process, could be obtained.This result well agreed with a diå erential curve of samplegravity obtained by the thermogravimetri c analysis. Itwas revealed that this system has a great reliability ofquantity. Furthermore, the present system is great eå ec-tive in the characterization of coal pyrolysis, because it isable to monitor continuously the reaction in situ up tohigh temperature (> 600 8C) other methods cannotexamine.

Using robotics to automate beryllium analysis byinductively coupled plasma-atomic emission spec-troscopy

Gerald L. DeVault, Greg G. Howard, Craig C. Hanzelka, Ed L.Churnetski and E. Ray Hinton, The Y-12 National SecurityComplex, Analytical Chemistry, 113C Union Valley Road, OakRidge, TN 37830-8045, USA

A robot has been developed to automate the samplepreparation of ® lter media for beryllium (Be) analysisby inductively coupled plasma-atomic emission spectro-scopy (ICP-AES). The robot increases the samplethroughput while reducing the potential exposure toanalytical technicians. Beryllium exposure can result in

a lung disease known as berylliosis, consequently work-place contamination must be continuously monitored.Beryllium has been used extensively in the nuclearindustry and can be found as a solid, a powder, or ina compound such as beryllium oxide. Typically, ® ltermedia is used for both air and surface monitoring,followed by labour intensive sample dissolution andanalysis by ICP-AES. In contrast, the robot automatesthe sample preparation and interfaces with the ICP-AES instrument resulting in complete automation of theanalysis. The ® lter samples are placed in 50-ml plasticcones labelled with a bar code. The robot transports thesample between stations where a number of functionsare performed, such as reading a barcode, ® lter dissolu-tion, addition of an internal standard, and dilution.After the sample preparation is complete, the robotsignals the ICP-AES instrument to start sample analy-sis. In 24 h, about 300 samples can be prepared,analysed and reported. Filters (n = 40) spiked with0.20 mg Be-nitrate were analysed and a mean of0.20 mg (§0.01 mg) was obtained. A detection limit of0.10 mg of Be/® lter has been demonstrated. Implementa-tion of the ICP-AES robot will increase the eæ ciency ofthe laboratory without reducing the quality of theanalytical results.

Flow-injection system for on-line olive oil emulsi-� cation and inductively coupled plasma-massspectrometry multi-elemental determinationsand in-situ emulsions stabilization for direct de-termination of iron and chromium in olive oil byelectrothermal AT

Juan R. Castillo, Maria S. Jimenez and Rosario Velarte,University of Zaragoza, Analytical Chemistry, Ciudad Universi-taria Scienca, Zaragoza, Spain

In this study we propose ® rst the on-line preparation ofemulsions by FI for multi-elemental determination inolive oil by ICP-MS. Besides another analytical interestof this study is to check the possibility of using aqueouscalibration solutions to quantify olive oil samples andcharacterize diå erent types of olive oils from diå erentSpanish areas Optimization of the chemical conditionsfor the on-line formation and stabilization of olive oilemulsions will be reported: type of emulsi® er, emulsi® erconcentration, sample amount, pH, ultrasonic stirringtime, salinity. In addition, optimization of FI variableswill be reported: amount of olive oil sample, ¯ ow rates,lengths and diameters of the diå erent tubing segmentsand ultrasonic stirring reactor. As a complementary workwe propose the determination of Fe and Cr in diå erenttypes of olive oil samples by GFAAS with direct oilemulsion formation by adding directly the sample andthe emulsi® er in water solution to the autosampler cup.The emulsion is formed by ultrasonic agitation with ahigh intensity ultrasonic processor used normally forslurries formation. Once the emulsion is formed, theautosampler injects the olive oil emulsion in the graphitefurnace and Fe and Cr are determined by AAS. Thismethod is very low time consuming and the samplepretreatment takes only 10 s. Diå erent parameters re-garding to the emulsion formation have been optimized:sample (olive oil) amount, emulsi® er concentration, time


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of agitation, ultrasonic power of agitation. In addition,parameters to do with the GFAAS determination havebeen studied: temperature programme and the use ofmodi® ers.

Castillo, J. R., Jimenez, M. S. and Ebdon, L. Journal of Analytical andAtomic Spectrometry, 14 (1999), 1515.

New on-line trace boron analyser for ultrapurewater

Richard D. Godec1, Paul Kosenka1 and Karla Dennis2, 1IonicsInstruments, Inc., Sievers New Product Development, 6060 SpineRoad, Boulder, CO 80301-3323, USA, 2Intel, Corporate ServicesFacility Tech, 2501 NW 229th Avenue, Hillsboro, OR 97124-5503, USA

We have developed a sensitive on-line analyser to meas-ure trace levels of boron in ultrapure water. This analyseris used in the semiconductor industry to control the levelsof boron in the ultrapure water used to make semicon-ductors. The analytical method will be described. Thedata from a performance study at an Intel semiconductorfactory will be reviewed. The study compared recovery ofboron standards from 0.030 to 2.0 ppb as boron inultrapure water using high-resolution ICP-MS and thenew on-line Boron Analyzer. The results indicate theboron standards ’ analysis data from the on-line BoronAnalyzer and the high-resolution ICP-MS are equiva-lent.

A screening method for polycyclic aromatic hy-drocarbons using hollow � bre membrane solventmicro-extraction

Stephanie King, Jill Meyer and Anthony R. J. Andrews, OhioUniversity, Chemistry and Biochemistry, 171 Clippenger Labs,Athens, OH 45701, USA

As regulatory agencies are faced with increasing work-loads, the development of a fast, low cost, reduced wastemethod to screen samples is crucial. A rising environ-mental concern is the contamination of water and soilwith polycyclic aromatic hydrocarbons (PAHs). Long-term exposure to PAHs has resulted in cataracts, kidneyand liver damage, reproductive diæ culties, and manydiå erent types of cancer. Sources of PAH contaminationinclude leakage from storage tank linings and burning ofoil and fossil fuels. Solvent Micro-Extraction (SME) is aneå ective method for analyte preconcentration providinglow limits of detection. However, in viscous solutions suchas soil, where rapid stirring improves extraction eæ -ciency, SME is often hindered by drop dislodgement.For this study we used hollow ® bre membrane solventmicro-extraction (HFMSME). A total of 8 ml of extrac-tion solvent was injected into a hollow, porous ® bre thatwas sealed on one end.

Automated on-line monitoring of VOC from thePAMS list

Philippe Verdeguer1,2 and Franck Amiet1, 1Chromato-Sud,R&D, 15 Route D’Artiguelongue, F-33240 Saint-Antoine,

France, 2Chromato-Sud, Department of Chemistry, 179 ChemistryBldg, Auburn, AL 36849-5312, USA

Volatile organic compounds (VOC) represent more than300 identi® ed compounds. Non-methane hydrocarbonsfrom two to 10 carbon atom number are the main part ofVOC in urban ambient air. Owing to their negativeimpact on human health, they are now considered are® rst priority pollutants. Benzene and 1,3-butadiene areknown to be highly carcinogenic. Moreover, they havebeen recognized to play, together with nitrogen oxides,an active part in tropospheric ozone formation. For thesereasons, governmental authorities now recommend thesurvey of individual VOC. In Europe, experts agreed ona list of 30 compounds, while in the USA 50 compoundshave been retained in the famous PAMS list. The air-mOzone rack oå ers the possibility of monitoring con-tinuously a wide range of ozone precursors down to pptlevels. The system is constituted of two gas chromato-graphs (4 U), with each unit being dedicated to theanalysis of a speci® c range of compounds: the Airmo-VOC C2± C5 allows the analysis of C2 to C5 VOCs, asthe AirmoVOC C6± C10 is dedicated to higher com-pounds. The two chromatographs can be integrated ina cabinet equipped with an H2 generator, a zero airgenerator and a permeation unit making a completeautomatic and stand alone system with no need ofexternal cylinder. Data on the stability of the system interms of retention time and peak areas as well as thelinearity of the response will be shown. The capabilities ofthe software as well as the possibility to performingremote control and calibration of the system will bedemonstrated. Finally, real life examples with identi® ca-tion of VOCs from the PAMS list will be presented. Thesample is stirred at 1350 rpm. After the optimized 8-minextraction time, 4 ml of the solvent was withdrawn anddirectly injected for analysis. Using an HP 6890 gaschromatograph with ¯ ame ionization detection, 17 PAHswere identi® ed based on retention time and furtherquanti® ed. Detection limits were calculated based on asignal-to-noise ratio of 3 and range from 0.132 to0.22 mg kg¡1 in soil. PAHs with low molecular weightsgave lower limits of detection because of their increasedsolubility in water. Those with higher molecular weightswere not extracted as eæ ciently. Calculated extractioneæ ciencies in soil varied from 1.8 to 8.2%. HFMSMEprovides a quick, simple, low cost screening method forenvironmental samples. Because the 8-min extraction canoccur during the 10-min GC run, ® ve samples can beanalysed per hour. Each extraction used one ® bre costingless than 1 pence and very small volumes of organicsolvent. No extensive ® ltering or pretreatment of thesample was necessary and the limited equipment re-quired would allow this technique to be used for on-siteanalysis.

An automated solid-phase micro-extractio n ofethyl carbamate in wines using gas chromatogra-phy/mass spectrometry

Eshwar Jagerdeo1 and Ingo Christ2, 1Department of Treasury,Bureau of Alcohol, Tobacco and Firearms, Department ofTreasury, 1401 Research Blvd, Rockville, MD 20850-3159,


206

USA, 2Leap Technologies, PO Box 969, Carrboro, NC 27510-0969, USA

Ethyl carbamate (EC), also known as urethane, is aknown carcinogen found in various food and alcoholbeverages. In fermented alcohol beverages, it is formedas a natural by-product . Currently, the US Food andDrug Administration (FDA) is doing the toxicologicalevaluation of EC with an aim of establishing maximumtolerance levels of this contaminant in food and bev-erages. Various methods have been published for thedetermination of EC in food and beverages. Thesemethods involve an extensive sample preparation pro-cedure with the potential for matrix interferences. Thisarticle discusses a method for the analysis of EC in winesusing an automated solid-phase extraction (SPME) anda gas chromatography coupled to a mass spectrometer inthe chemical ionization mode. In this method, variousconditions and parameters of the ® bres and samplepreparation are optimized for the quantitation of EC inwines. The use of a dual-arm autosampler helps in theautomation of the sample preparation. Furthermore, theSPME procedure uses a heated magnet-mixing chamberto reduce the sample extraction time. This methodeliminates the need for extensive sample preparationand aå ords EC determination and con® rmation in thelow ppb. This procedure is quick, and aå ords excellentquantitation and reproducibility.

Automated on-line monitoring of volatile sulfurcompounds for environmental and industrial ap-plications

Philippe Verdeguer1,2 and Franck Amiet1, 1Chromato-Sud,R&D, 15 Route D’Artiguelongue, F-33240 Saint-Antoine,France, 2Chromato-Sud, Department of Chemistry, 179 ChemistryBldg, Auburn, AL 36849-5312, USA

Because of their impacts on environment and humanhealth, the analysis of volatile sulfur compounds hasbecome a necessity. This paper presents some applica-tions of the new generation of Airmomedor analyser.Based on gas chromatography with a speci® c electro-chemical detector for sulfur compounds, this on-lineanalyser allows the survey of H2S, COS, mercaptansand sul® des at the ppb level. It can be integrated in acabinet with a zero air generator, a permeation unitmaking a complete stand-alone unit with no need ofexternal cylinder. It can operate in a fully automaticmode including remote control capabilities and auto-matic validation. Datas on stability and linearity of theresponse will be shown. Diå erent applications will bepresented including fermentation process control, qualityof CO2 or other gas process, waste water plant treatmentprocess control, continuous emission monitoring and airmonitoring stations.

The combination of an automated trappingsystem with a portable GC

Wolf Muenchmeyer and Andreas Walte, WMA Airsense,Hagenower Str. 73, Schwerin, MV D-19061, Germany

A new trapping system was developed in order tocombine it with a portable GC. The detection system

consists of a micro-GC using a thermal conductivitydetector. For some applications the detection limit ofabout 1± 10 ppm for the detector is too high. Sometimesthere is also a higher selectivity needed. With an externalenrichment unit, it is possible to lower the detection limitby one or two dimensions, depending on the samplingtime and the retention capabilities of the adsorptivematerial. By choosing an optimal adsorptive materialwith the corresponding sampling temperature, a selectiveenrichment is also possible. Disturbing solvent peaks orother main peaks due to humidity can be eliminated orminimized. After sampling, the adsorption tubes arethermally desorbed and analysed. Applications showingthe reproducibility and the improved detection limits willbe shown.

Improvement of sample carryover in the auto-sampler of HPLC

Nobuyuki Tatsumi, Yoshiaki Maeda, Tsuyoshi Morikawa,Yoshiaki Aso, Shuzo Maruyama and Teruhisa Ueda, ShimadzuCorporation, 1 Nishinokyo-Kuwabaracho Nak, Kyoto 604-8511,Japan

Sample carryover becomes one of the most importantperformance to evaluate the HPLC system especially forthose who uses HPLC in high-sensitivity analysis. As theMSMS detection is getting popular in the high-sensitivityHPLC analysis such as pharmacokineti c analysis, itappears that the carryover in the HPLC system is some-times too much to be ignored because the sensitivity ofthe MSMS is much higher compared with the UV-VISdetector and the more the sensitivity is, the higher thecarryover is observed. There are several causes of thecarryover and it depends on the chemical characteristicsof samples, ¯ ow line con® guration of the autosampler,washing liquid and procedure of the autosampler, etc.However the two major cause of carry over are a ionicinteraction between the sample and the metallic parts ofa ¯ ow line such as a sample-sucking needle and ahydrophobic interaction between sample and polymericmaterial in a ¯ ow line such as Vespel or PEEK used for aswitching valve of the autosampler or other plumbing. Inthis study, several types of sample-sucking needle havebeen compared when concerned with the carryoverperformance with both ionic and non-ionic substances.In addition, some materials used for the plumbing suchas SS or PEEK tubing have been examined for chemicalinteractions to eliminate the carryover of HPLC system.

Liquid chromatography/mass spectrometry withon-line photochemical derivatization of selectedindole compounds

Beata A. Musial, Abdulqawi Numan and Neil Danielson,Miami University, Chemistry and Biochemistry, 112 HughesHall, Oxford, OH 45056, USA

Over the years, mass spectrometry (MS) has been provento be the method of choice for obtaining importantmolecular ion information of many compounds. The highsensitivity and reliability of MS have attracted manyscientists from various ® elds. The identi® cation of isomersand closely related compounds is an existing problem


207

with mass spectrometry especially if tandem mass spec-trometry (MS/MS) is not available. Previously, we foundthat the combination of on-line photoderivatization and¯ ow injection analysis-mass spectrometry (FIA/MS)can be successfully applied for the identi® cation of aro-matic compounds such as sulfonamides that have verysimilar structures. A photoreactor with the dimensionof 2.5 m £ 0.25 nm and a volume of 0.12 ml was used.Without photolysis sulfonamides tend to follow a com-mon fragmentation pathway. Upon photolysis, however,each sulfa drug showed a unique mass spectrum thatmakes its identi® cation much simpler. In the presentwork, the on-line photochemistry and MS of indoles willbe addressed. Indole derivatives such as serotonin, tryp-

tamin, tryptophan , melatonin and indole-3-acetic acidexhibit a UV absorbance in the region 250± 290 nm,which makes their identi® cation with UV-Vis diæ cult.The photolysis of indoles is of interest since some can beconnected to the photodegradation of protein and someare of interest as antioxidants . For this work, Te ontubing reactor (4 m £ 0.1 mm i.d., 0.4 mm o.d.) will betested. We have found that the volume of the photo-reactor has a major role on the degree of the photo-transformation of the indole compounds. Additionally,the nature of the mobile phase as well as the ¯ ow ratesin¯ uence the photolysis products of the indoles. Finally,the LC separation of a mixture of indoles with on-linephotolysis and MS will be addressed.


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