Automatic The 1991 International Symposium on Laboratory ...

Journal of Automatic Chemistry, Vol. 13, No. 6 (November-December 1991), pp. 243-266

The 1991 International Symposium onLaboratory Automation and Robotics

Nearly 380participants gathered in Bostonfor the ninth meeting heldfrom 20 to 23 October 1991. They camefrom many parts ofthe UnitedStates andfourteen other countries. Theformatfor the conference includedplenary lectures, other oralpresentations, posterpapers and a visit tothefacilities ofthe meeting hosts, Zymark Corporation in Hopkinton, Massachusetts. Following the keynote speakers on Monday morning,there were sessions on Pharmaceutical analysis and advanced technology. Theformer continued into the afternoon in parallel with session onIntelligent automation and microplates. Posterpresentations completed the afternoon schedule. Theprogramme on Tuesdayfollowed a similarpattern. Morning sessions on Managing laboratory automation and Environmental ran in parallel. The latter continued on in the afternoonalong with a session on Biology/biotechnology. Poster sessions again completed the afternoon programme. General papers were presented onWednesday in parallel with a session on chromatography sample preparation.The plenary programmefeatured two speakersfrom the pharmaceutical industry; David B. Weinstein of the Sandoz Research Institute andBernd C. Schade of Miles Incorporated. Their topics were, respectively, Jumping into the 20th century before it’s too late; is laboratoryrobotics still in its infancy.), and ’Quality control in theyear 2000’. It is interesting that while pharmaceuticalfirms early on recognised thecontribution robotics could make to laboratory automation, the#focus is on the future.Five of the six speakers in the session on Managing laboratory automation were also from the pharmaceutical industry. A theme that ranthrough the presentationsfrom thepharmaceutical industry is the needfor support ofsenior management in the development ofroboticfacilitiesand thefact that this commitment must befor the long haul. Ofcourse, the bottom line is better quality products and increasedproductivity.Another feature of the meeting is the presentation of the Pioneers in Laboratory Robotics Awards. The recipients for thisyear were:

Timothy Lincoln (Xerox Corporation)Mark Arnold (Bristol-Meyers Squibb)Frank Dias (WMI Environmental Laboratories Inc)Lisa Martin (ICI Pharmaceuticals)Richard Young (RW Johnson Pharmaceutical Research Institute)Katherine Ku (Schering Plough)Mike Metzger (The Goodyear Tire & Rubber Company)Stephen Conder (Bristol-Myers Squibb)Kevin J. Halloran (Carter-Wallace Inc)

The abstracts of thefull programme are included in this issue 0fJournal of Automatic Chemistry by kind permission of the ZymarkCorporation and the papers will be available in early 1992 as ’Advances in Laboratory Automation and Robotics’, Vol. VIII.This meeting is well worth attending. The tenth symposium will be heldfrom 25 to 28 October 1992 and information is availablefrom SharonCorreia, Zymark Corporation, Zymark Center, Hopkinton, Massachusetts 01748, USA. Tel.: 508 435 9500. Fax: 508 435 3439.

DAVID J. CURRANAmerican Editor

Abstracts from the 1991 International Sym-posium on Laboratory Automation and Robotics

KEYNOTES

Jumping into the 20th century before it’s too late: islaboratory robotics still in its infancy?

David B. Weinstein and Dennis S. France (Sandoz ResearchInstitute, East Hanover, NJ 07936, USA)Successful management oflaboratory robotic automationprograms in the area of research and drug discovery inthe pharmaceutical industry may perhaps be best com-pared to a chef preparing the perfect sauce. All theingredients must be available at the same time and be ofhighest quality for the right price. However, if compo-nents are not added in the right quantities, and in the

proper order, no amount of whipping together by theproduct champion will create the best product. In thepast, managerial scepticism surrounding useful imple-mentation of cost-effective, high throughput roboticsystems often placed these ’modern toys’ at low prioritiesfor research development laboratories. Management nowrecognizes the unique contributions of robotics in theresearch environment. Although the scientific directormust still play the role of product champion, newquestions are being proposed and new commitments arebeing made to bring the potential of robotic automationto every laboratory where repetitive functions can benefitfrom new applications. Research laboratory directorshave become both the key ingredient, as well as the rate-limiting determinant in the development of new appli-cations. Having fulfilled the promise of robotic automa-tion to release talented personnel, the challenge now is forthe ’end users’, the bench scientists, to be provided withopportunities to critically invest time and effort requiredfor future applications and new career functions.

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The 1991 International Symposium on Laboratory Automation and Robotics: Abstracts

Quality control in the Year 2000

Bernd C. Schade (Miles, Inc., Consumer Healthcare Division,Elkhart, IN 46515, USA)

’Just-In-Time’ production is a prerequisite for a copy tomeet the challenges ofcompetition. Manufacturing cycleshave been so successfully optimized, that release time hasbecome a significant factor. A vision for a major QCcontribution to profitability in this decade seems to be theJust-In-Time-Release. Benefits will exceed cost savingsfor lower inventory, because problems are detected asthey occur. To meet this goal, the requirements forautomated instruments will change significantly.Quality, reliability, durability, and ruggedness of auto-mated systems will be crucial factors for success.

PHARMACEUTICAL ANALYSIS

Better chemistry through robotics: an automatedsystem for process optimization

Susan D. Boettger (Bristol-Myers Squibb Company, Syracuse,NY, USA)

Rapid development of an economical process for manu-facture of the active ingredient plays a key role in thesuccessful launch ofa new pharmaceutical. The evolutionfrom a multistep laboratory synthesis of milligramamounts to the streamlined manufacture of multikilo-gram quantities is typically a slow and labour-intensiveprocess. The Bristol-Meyers Squibb Process Optimiz-ation Robotic System was designed to shorten thisprocess by increasing the quantity and quality ofreactions investigated. In addition, automated reactionsetup and monitoring frees the development chemist formore creative activities.

This author described the three-phase development oftheProcess Optimization Robotic System. In Phase 1,automation of a 2 h, room temperature reaction wasachieved via hardware and software modifications of acustom Zymark PyTechnology system. The second phaseinvolved installation of a low temperature cooling systemand control of reaction temperatures from -60C to125 C via EasyLab programming. Phase 3 comprisedoptimization of a palladium-catalysed reaction by vary-ing concentration, temperature and the amount and typeof catalyst and additives. The ability to quantitativelyassay multiple reactions during overnight runs resulted inreduction of the reaction time from 20 to 2 h and rapididentification of optimal reagents and temperature.

Robot automation of routine assays in the stabilitytesting of solid dosage forms

Steven Conder (Bristol-Myers Squibb, New Brunswick, NJ,USA)The increasing demands of stability testing of dosageforms during drug development have created an immensesample load for routine assays such as in vitro dissolution

testing, composite and single tablet High PerformanceLiquid Chromatographic (HPLC) assays, and KarlFischer moisture analysis. The productive capacity of thelaboratory which uses manual or semi-automated meth-ods is easily overwhelmed by the volume of samplesgenerated by these studies. The need for fast, accurate,and flexible automation capable of solving the variety ofanalytical problems facing the laboratories involved withstability testing is necessary to speed drug development.The New Brunswick Arialytical Research & Develop-ment of Bristol-Myers Squibb has designed a laboratorydedicated to robot automation of the routine time-consuming assays involved with stability testing. Thelaboratory was designed to contain up to nine robots.Currently six Zymark robots and a Source for Automa-tion (SFA) tablet extraction workstation are operational.The Zymark system types include four dissolution testingrobots, one HPLC assay robot and a dual-function robotfor Karl Fischer moisture determinations and tabletassays by wet-homogenization. This paper discussesseveral examples of successful assays performed by theserobots, including the problems of system validation andsample/data management. The laboratory was successfulin completing over 20 000 tablet assays in its first twoyears of operation.

Robotic moisture determination of solids in thepharmaceutical industry

Edward G. Kanczewski, Marie Skrilec and Robert E. Daly(Parke-Davis Pharmaceutical Research Division, Warner-Lam-bert Company, Morris Plains, NJ, USA)Accurate moisture determination of formulation ex-cipients and active ingredients is a major concern inthe pharmaceutical industry. Unassisted Karl FischerTitrations are performed using a Zymark robotic systemand appropriate Metrohm equipment. Samples having arange of moistures from approximately 0"5% to 10% areanalysed in triplicate to insure precise and accurateresults. User-friendly screens prompt the operator andallow the novice to use this system. Restricted access toZymark software minimizes the curious from modifyingsystem operating software.

’On-line’ monitoring of a pharmaceutical tabletpress using a customized laboratory workstation

D. Jeff Roberts, Michael A. Shirley and Frederick K. Bangert(Bristol-Myers Products Division of Bristol-Myers SquibbCompany, 9707 Chapel Hill Road, P.O. Box 300, Morrisville,NC 27560, USA)The method for determining Uniformity of Dosage(Content Uniformity) for solid dosage form pharma-ceutical products, as defined in USP XXII, was auto-mated using a customized Tablet Assay Workstation.The workstation performs all of the sample and glasswaremanipulation including HPLC analysis, cleaning anddisposal. The system handles all sample transfers, as wellas the addition of diluting solutions and internalstandards. Sample analysis and quantitation is per-formed by an on-line HPLC injection and data collection

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system. The sample transfers and diluting solutionadditions are made with calibrated syringes, peristalticpumps and automated pipetting hands, and the transfersare verified gravimetrically. In addition, this system hasthe ability to wash and dry the sample-processingcontainers for re-use by the system.

The goal is to incorporate the on-line sampling andphysical testing (weight/thickness/hardness) capabilityof an automated tablet press with the capabilities of theTablet Assay Workstation to generate real-time data for"monitoring the performance of the tableting process.Ultimately, batch production and testing will be droppedin favour of continuous production and testing. Theprimary benefit of this system merger will be an overallincrease in plant productivity through improvement ofQuality Assurance throughput time, without compromis-ing product integrity or process control, whilst meetingUSP XXII Uniformity of Dosage Units requirements.

Automated performance of particulate matter test-ing for parenteral products

Thomas A. Barber, Christine Pavek-Hicks andJohn G. Williams(Baxter Healthcare Corporation, Round Lake, IL, USA,)The enumeration of particulate matter in parenteralproducts provides a critical assessment of process cleanli-ness and product quality. The United States Pharmacopeia(USP) defines limits for allowable numbers of particles inboth large volume parenteral (LVP) solutions and smallvolume injections (SVI). It also describes a test methodfor particulate matter based on light obsuration particlecounters; this method is extensively applied by pharma-ceutical manufacturers as a quality assurance and releaseassay.

The goal ofthe automation project described in this paperwas the elimination of the manual component of theinstrumental particulate matter test routinely performedon large numbers of samples. This was achieved bycombining a Zymate robot with a HIAC particle counter.The performance criteria for the system include: (1)unattended operation; (2) fail-safe measures preventingincorrect unit identification; and (3), of extreme impor-tance, logic which allows scrutiny of counts collected forartifacts before data is sent to a database. Due to thetransfer of this work to a Zymate System, the techniciansresponsible for a test now spend only the time necessaryto set the system up and load units to be tested.Additional benefits include improved precision of dataand the ability to test outside of normal working hours.

Automation of dissolution testing using robotics andnetwork-based data management

R. Foester, L. Nemec andJ. Rosen (Schering-Plough, Kenilworth,NJ, USA)

The requirements for dissolution testing have increaseddramatically in recent years. At Schering-Plough Phar-maceutical Research, dissolution testing has beenassigned to a specialized group which supports all solid

dosage formulation research and development/improve-ment preceding product approval. Only through exten-sive automation and electronic data handling couldadequate support and productivity be provided.

Dissolution testing involves the following four categoriesof laboratory work:

(1) Dissolution test set-up and clean-up.(2) Dissolution run (basket, paddle) with sample

collection.

(3) Quantitative analysis (UV, HPLC).(4) Sample data management.

The majority ofsamples are tested semi-automatically onthe equipment described below. The semi-automaticapproach cannot handle the first task: test set-up(preparation of dissolution media, filling of vessels) andthe final vessel clean-up. Integration of two Zymarkdissolution robots into the system permits automaticvessel filling and clean-up for the four dissolutionapparatuses serviced by robots.

Samplers and timers were designed for the semi-automatic sample aliquot collection. Trays holdingsample containers are transferred from the samplers tothe sippers attached to the spectrophotometer. Anelectronically controlled valve connected to housevacuum is used to sip aliquots through a flow-through cellon the spectrophotometer. No sample dilutions arenecessary- the absorbance of aliquots is kept withinquantitative limits by the selection of spectrophotometricflow cells with suitable optical path length.

Quantitative analysis of dissolution sample aliquots canbe performed either on-line, during the actual dissolutionruns (several commercial systems are available) or afteraliquots have been collected in individual sample con-tainers. For the semi-automatic mode we chose the latterapproach since one spectrophotometer can easily runassays of samples collected on four or more dissolutionapparatuses. The dissolution robots can operate eitheron-line or off-line, depending on the types of samplepreparation and quantitation required (additional fil-tration, HPLC assays require off-line sample collection).

The data management portion of the system (custom-designed dissolution LIMS) has been instrumental inmaking highly productive and reliable data reportingpossible. Samples are run using stored descriptors whichare decoded by the software to select the appropriateassay method. Seven identically equipped IBM PC-basedworkstations are on-line and connected by a Local AreaNetwork residing on a server equipped with a largestorage capacity an automatic tape back-up assuringpermanent and secure storage of all primary spectral anddissolution data.

The automation ofdissolution testing at Schering-Ploughrequired careful optimization ofresources and a consider-able amount of hardware and software design. Thepresent approach is just one of many alternativespossible, all of them requiring extensive customization,since a viable, high-volume commercial system is notreadily available.

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Automated HPLC analysis using a Zymark RoboticSystem

K. K. Chow and Aurobindo Nair (Glaxochem, Ltd) and P. C.Thiak and Teoh Sek Hong (Microstate, Singapore)

This paper presented the approach taken in automating a

high volume, labour intensive operation. The applicationis in the quality control laboratory of Glaxochem (Pte)Ltd in Singapore. It involves the HPLC assay of the H2-antagonist, Ranitidine HCL, which is the active ingredi-ent in the prescription drug, Zantac.

The manual HPLC assay is a tedious and time-consuming operation. This analytical routine has beenautomated by interfacing a Zymate II Robot, with theappropriate PySections, to a HPLC system. The robothandles all aspects ofsample preparation and subsequentinjection onto the HPLC system. This configuration hasdramatically improved sample turnaround time andenhanced the accuracy and precision of the analyticalresults.

Aspects of the application programme and procedureundertaken in validating the complete system werediscussed. Problems encountered during validation andthe steps taken to overcome them were highlighted; andthe utility of gravimetric laboratory techniques in,improving accuracy and precision of analytical resultswas also explained.

ADVANCED TECHNOLOGY

Computer validation: how much is enough?

James E. Curley (Pfizer, Inc., Groton, CT, USA)The author discussed the type of computer system thatneed validation from the managerial point of view ofbalancing costs and benefits. The requirements of thosedemanding validation must be taken into account inplanning the experiments and documentation for valida-tion. Approaches to developing test protocols, test cases,change control and documentation were described.

This presentation discussed the new technologies that areavailable and their possible integration into automatedsystems. Automation challenges for the 1990s were alsodiscussed.

Hands-free GC-Zymate to chromatography datasystem via Ethernet

C.J. Deakin and M.J. Crook (BP Research Centre, Sunbury-on-Thames, UK)

Many laboratory procedures can be automated to givesignificant improvements in accuracy, precision, speed ofanalysis and sample throughput. This automation invar-iably involves a variety of software controlled equipmentwhich is rarely fully integrated into a completelyautomated laboratory.

The analysis of samples by GC Simulated Distillationinvolves extensive sample preparation, GC analysis, datacollection and post analysis calculation. The completeanalysis has been fully integrated through the develop-ment of in-house software to interface a Zymate PyTech-nology automated sample preparation system with theVAX/VMS based VG DataSystem Chromatographydata system (Multichrom). The robot controller PC hasbeen connected to the site computer network via Path-works for DOS software and the DECnet transportprotocol. Using this approach to the transfer of sampleparameters between devices has eliminated any manualre-entry or computational reformatting of the data. TheVAX/VMS component of the software is installed as aDECnet object on the VAX node and is activated fromthe PC. Subsequent communication is via non-transpar-ent task-to-task link between the two processors.

The advantages of this integrated approach were dis-cussed, with special emphasis on the communicationslink between microprocessor based equipment fromseveral vendors.

New analytical technologies and the challenge toautomation

W. Jeffrey Hurst and Robert A. Martin, Jr. (Hershey FoodsTechnical Center, Hershey, PA, USA)

Many discussions of laboratory robotics and automationconcentrate on the use of robotics in sample preparation,but fail to discuss their integration into the overallanalytical method. In recent years there have been rapidadvances in several new analytical technologies. Two ofthe most intriguing developments have been SupercriticalFluid Extraction (SFE) and Capillary Electrophoresis(CE). Each of these technologies has its own set ofuniqueproblems. Additionally each has limited automationcapabilities and therefore presents challenges to thedevelopment of next generation automated systems.

INTELLIGENT AUTOMATION

The analytical director project

Thomas L. Isenhour (Department of Chemistry, Kansas StateUniversity, Manhattan, KS 66502, USA)

The goal of the Analytical Director project is to build a

system that can design, test, modify and implement itsown analytical procedures. The Analytical Director is acombination of commercial robotic and other laboratoryequipment with computer based artificial intelligence.The prototype system is now able to design an entireanalytical procedure in a high level language workingfrom an icon driven windowed display and have the robotlaboratory execute that procedure without humanintervention.

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Flexible, intelligent systems for automated chemicalanalysis

Frank A. Settle, Jr. (Department of Chemistry, U.S. Air ForceAcademy, USAF Academy, CO, USA), H. M. Kingston(Department ofChemistry, Duquesne University, Pittsburgh, PA,USA) and Michael Pleva (Department ofChemistry, Washingtonand Lee University, Lexington, VA, USA)Commercial expert system shells, spreadsheet and data-base programs can be combined with automated labora-tory equipment to form flexible, intelligent systems. Aftera brief introduction, three examples were presented. Thefirst system integrated an expert system shell, VP-Expert,with a Zymark System V robot to perform qualitativeanalysis for selected metal cations. The second systemcurrently under development, combines VP-Expert,Lotus 123 and an experimental design program with theZymate System and a Milton Roy spectrophotometer forthe design and implementation of.colorimetric phosphatedeterminations. The final system involves the PC-Plusshell and dBase III linked to a Zymark robot and a CEMmicrowave unit to perform dissolution of a variety ofsample matrices.

automated systems. The encapsulation system combinesa microwave procedure generation, sample log-in, cali-bration, and microwave control program, and variousversions of system hardware control software. Thehardware consists of an integrated Zymark robot withperipheral equipment, System V controller, and a PCclone to control the automated microwave equipment.The microwave procedure generation program creates aprocedure file that includes information required for a

completely manual operation or for control of a fullyautomated system. The encapsulation of the method intoa computer file is the basis for procedure transfer betweenthese automated digestion systems. The two EPA sitesare currently running routine samples using this methodof encapsulation and rely on calibration to transfer themicrowave conditions.

This example was used to discuss these specific methods,as well as the general topic of encapsulating analyticalprocedures and their electronic transfer to automatedsystems. It may soon be possible to down-load analyticalchemical methods in the same way programs aretransferred today.

Encapsulation and transfer of standard methodsusing automated equipment

H. M. Kingston, Peter Walter (Duquesne University, Pittsburgh,PA, USA), and Frank A. Settle, Jr. (Department of Chemistry,U.S. Air Force Academy, USAF Academy, CO, USA)Microwave sample preparation is gaining a wide degreeof acceptance within the analytical community. TheEnvironmental Protection Agency (EPA) is in the processof approving two microwave dissolution methods fortrace elemental analysis (Methods 3015 for aqueoussamples and 3051 for soils, sediment, sludges, and oils).Required procedures such as these provide solid rationaltbr automation and new challenges for accurate methodstransfer. The nature ofstandard procedures requires thatthey be readily transferable and reproducible betweenlaboratories. Microwave sample preparation offers twoviable methods for transfer of microwave conditions:equipment calibration and feedback control.

Calibration of microwave equipment permits transfer ofprocedures between various models and manufacturersby replicating the microwave field flux of the equipment.Feedback control of microwave equipment is based onreal-time measurement of temperature and is also aviable way of transferring the test conditions.

Until recently, procedures were transferred betweenanalysts using mainly manual equipment. With thedevelopment of automated equipment, a new aspect ofmethod transfer must be addressed- the electronictransfer of analytical chemical procedures. Three auto-mated microwave sample preparation systems have beendesigned and are located at EPA sites in Las Vegas,Nevada and Seattle, Washington, and at the NationalInstitute of Standards and Technology (NIST, Gaithers-burg, MD). Methods to capture the environmentalprocedures are being evaluated and tested with these

Objected-oriented data handling system for an auto-mated chemistry laboratory

P. A. Medvick, S. M. Mniszewski and T.J. Beugelsdijk (LosAlamos National Laboratory, Los Alamos, NM, USA)

The environmental-remediation efforts at DOE com-plexes require characterizing problems at each site beforetaking clean-up action. Characterization will require thechemical analysis of millions of samples at a significantcost. Automation of the required chemical analysesmethods provides a cost-effective solution. Five of theDepartment of Energy (DOE) national laboratories, LosAlamos National Laboratory (LANL), Pacific NorthwestLaboratory (PNL), Sandia National Laboratories(SNL), Idaho National Engineering Laboratory (INEL),and Lawrence Livermore National Laboratory (LLNL)have joined together to automate the environmentalchemistry laboratory. An object-oriented approach wasdeemed necessary to allow for modularization, maintain-ability, reusability, and flexibility of the software andhardware.

Each chemical analysis method is implemented as aStandard Analysis Method (SAM). A SAM is, in essence,a ’black box’ into which a sample enters at one end andchemical or physical data ’exists’ at the other. A SAM iscomposed of a set of Standard Laboratory Modules(SLMs) implementing sample preparation operations(such as weighing, grinding etc.), analytical instrumen-tation operations (such as measurement control etc.), anddata interpretation operations such as (data reductionand documentation). Each of the SLMs are modular,standardized, and highly automated.

The software system is being developed in C+ + buildingupon existing code developed by SNL for a Waste StorageTank Generic Controller. The C++ based, object-oriented database, ONTOS by Ontologics, is being used

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to store the knowledge for operating the automatedlaboratory as well as the information collected from thesample analysis. ONTOS provides a graphical tool fordatabase construction, C++ library routines for pro-grammatic interaction, ONTOS SQL for queries withinapplication programs, and a tool for visually andinteractively creating graphical user interfaces for data-base access.

Database objects created were selected for quality con-trol, hardware set-up and control information, and datainformation. The ADISS constructs by R. Lysakowskiwere used as a basis for the initial database design, whichwas subsequently modified to conform with the SAMsystem requirements. The following major objects wereidentified for this database: administrative, sample,aliquot, physical component, SLM, SAM, supplies,operation-report, status, calibration-results, raw-data,and processed-data. This knowledge, its structure, andhow it is used to drive the system were further explored inthe paper.

MICROPLATES

Fluid handling evaluations for microplate assays

Gerald Hahn (Beckton-Dickinson Advanced Diagnostics, Balti-more, MD, USA)

One of the most important aspects of an automatedmicroplate system are the fluid-handling components.These are usually stepper motor driven syringes operatedthrough a control interface. The accuracy and limitationsof these fluid handling instruments will be discussed. TheZymate modules and the Tecan 505 system werecompared. Improved user modules and programmingsuggestions were also discussed.

replacing fresh microtube racks into the fraction collec-tors, the robot performs a number oflipid and apoproteinELISAs on the fractions and then stores the fractions forfurther analysis. Despite the increased throughput of 96complete lipid (cholesterol, phospholipid, and triglycer-ides) and apoprotein (apo A-I, B and E) profiles in a 24-hour period, the robot arm was still idle nearly 70%during a robotic run. This idle robot time has beenexploited by splicing multitasking clinical chemistryanalyses previously run by a different robot into the toplevel FPLC application program. Two major innovationshave made this possible. Firstly, liquid handling pro-grams involving the multipipette hand have been stream-lined and optimized to allow variable volume aliquotingof serum samples into various replicate assays. Secondly,we have utilized the force feedback potential of thegripper hand to simply look for sample plates during therobot’s idle time while serum samples are fractionatingand collecting. Analyses are initiated when the robotperceives that the samples have been provided. Prelimi-nary data will be presented regarding both the ZymateXP arm on this system and novel applications by theother robot, having been freed up by the FPLC robot, ofhigh throughput cell-based drug discovery screens.

Modifications of robotic enzyme-linked immunosor-bent assay (ELISA) system for rodent serology toenhance capacity, throughput and sensitivity

William R. Shek, Paul A. Oskar and Mark A. Cerra (CharlesRiver Laboratories, Wilmington, MA, USA)Adventitious viral and mycoplasmal infections of labora-tory rodents can cause changes that alter researchfindings. Some viruses indigenous to rodents are zoonoticagents capable ofcausing disease in humans. Rodents aretherefore routinely monitored for viruses by assaying forspecific antibodies formed as part ofthe immune responseto infection.

The microplate robotic system as a complex bio-chemical and immunochemical detector for highthroughput chromatographic analysis of serumlipoproteins

Dennis S. France, Naved A. Surve, Robert E. Quinby, Mary E.Russell, Mary Kae Murdoch, Kathy Ramos, James R. Paterniti,Jr. and David B. Weinstein (Department ofLipid & LipoproteinMetabolism, Sandoz Research Institute, East Hanover, NJ 07936,USA)

While robotic systems have been frequently interfaced toHPLC systems, they usually play the role of automatedsample preparation prior to chromatographic analysis.These roles have been reversed in a system in which aZymate II Microplate Management System serves anHPLC system in a variety of functions. The prototype ofthis system has been previously described with onePharmacia Superose-6 FPLC column and one autoinjec-tor. The pivotal components of the current system arefour FPLC columns, two Gilson 203 fraction collectors,two Waters WISP autoinjectors, and two Waters WAVSautomated valve switching stations. In addition to

During the past year, the authors have switched from a

partially automated ELISA procedure to one that is fullyautomated, using the Zymate II Microplate Manage-ment System. The results of the evaluation of the roboticsystem and comparison of its sensitivity and specificity tothose of the non-robotic procedure were recently pub-lished (Cerra et al., LRA 2:119-131, 1990). In all aspects,including accuracy and precision of pipetting stations,and assay sensitivity and specificity, the fully automatedrobotic system compared favourably with the partiallyautomated ELISA.

Since that publication, modifications have been made tothe robotic system to further enhance its performance.The Zymate II controller was replaced with a System Vcontroller which (in contrast to the former) is able to readand write ASCII files. With the Zymate II controller,essential array variables were sent from a remote PC-XTto the robot via a Z845 interface. With the System Vcontroller, this slow step has been eliminated. Instead,the array variables, written to the files by the remotecomputer, are read directly from the controller disk drive.This is done at the beginning of each assay cycle so that a

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run can be started before all sample and test plates havebeen loaded. As a consequence, robot runs can be startedearlier in the day, thereby increasing the actual dailythroughput. The standard Zymark microplate storagerack holds 15 microtitre plates. These have been modifiedto hold an additional 5 plates per rack. By adding a 6thtest plate storage rack, we have increased the maximumnumber of test plates per run from 75 to 120. Anadditional 10-slot incubator has been placed on the robottable to double the test plates processed per assay cyclefrom 10 to 20 and the incubation period per step from 20to 40 min. This longer incubation has increased assaysensitivity. Other modifications that have improvedsystem performance and reliability were also discussed.

Automated affinity column chromatography system

Kirk Andree, Dana Branham, Sandy Marshall (HamiltonCompany, Reno, NV, USA), Martin. Gross (Isolab, Inc., Akron,OH, USA) and Robin Felder, Ph.D. (University of Virginia,Charlottesville, VA, USA)The automation of assay processes can make significantcontributions to throughput and assay precision, whilstreducing labour costs in most laboratories. Roboticpipettors have traditionally been used exclusively for theautomation of the pipetting in clinical pharmaceuticaland biotech laboratories. The system presented appliedthe robotic pipettor not only to pipetting, but also to thefull automation of an affinity chromatography methodbased on a gravity feed, disposable column. Theapproach using the (Glyc-affin) GHb (Isolab, Inc.)assay for total glycated haemoglobin in whole blood wasdescribed.

A Microlab 2200 robotic pipettor (Hamilton Company)was equipped with special hardware which automates thesteps involved with fractionating glycated haemoglobinfrom non-glycated haemoglobin. The manual procedureis labour intensive and can be error prone; it requiresseveral pipetting steps, extended column drain periods,mixing, dilutions, and critical timing steps. As thenumber of samples and columns to be processedincreases, the robotic system remains consistent, wherethe possibility for procedural error increases with themanual approach.

The automated system limits manual intervention toapproximately 30 min for assay set-up to load samples,columns fraction collection tubes, microplates and re-agents. After set-up the technologist can walk away forthe duration of the procedure (22’5 h). The roboticpipettor takes the whole blood sample and processes itthrough to the final fractions in microplates. Assay CVsare reduced from 7-10% manually to 2-3% withmicrolab Glyc-Affin, due to precision liquid handling,timing and column, manipulations. Total operator timeper 96 samples is 30 min with the automated systemversus 3 h manually. Data were presented comparing theassay performance and operator time for the manualmethod and the automated system. Alternative appli-cations, including solid phase extraction and filtrationwere explored and presented.

MANAGING LABORATORY AUTOMATION

The laboratory of the 90s planning for totalautomation

Linda A. Brunner (Ciba-Geigy Corporation, DevelopmentDepartment, Pharmaceutical Division, Ardsley, NY, USA)The analytical laboratory of the 1990s must be able tomeet and accommodate the rapid evolution of modern-day technology. One such area is laboratory automation.Total automation may be seen as the coupling ofcomputerized sample tracking, electronic documentationand data reduction with automated sample handling,preparation and analysis, resulting in a complete analyti-cal procedure with minimal human involvement.

This is a goal that many of us, in various industriesemploying analytical technology, are working towards aswe enter into the 90s. However, the requirements mayvary from one laboratory offacility to another. Therefore,the automation has to be flexible enough to cover a widerange of applications, and yet fit into specific nichesdepending on individual needs.

Total automation must be planned for, well in advance, ifthe endeavour is to be a success. Considerations must bemade for available space, laboratory layout, properequipment, and the availability and access to necessaryutilities. Adequate training and experience of the person-nel working with the technology must be ensured. Inaddition, responsibilities of installation, programming,maintenance and operation have to be addressed. Propertime management and the efficient implementation anduse of total automation are also crucial to successfuloperations.

The author’s insights into laboratory organization andrequirements, as well as management issues that must befaced when automating laboratory procedures werediscussed.

Measuring the effects of laboratory automation

Gerry Fitzgerald and ,Jim Swanson (SmithKline BeechamPharmaceuticals, Research & Development Division, King ofPrussia, PA 19406, USA)

A variety of scientific and management motivations leadto the automation of laboratory systems. Assessing theimpact that automation has had on the organization is apart of maintaining these systems. SmithKline BeechamR&D is using several different types of measurement andmany different tools for measuring the effects of auto-mated laboratory systems. These metrics show howautomated laboratory systems are affecting the workflowand information flow in each laboratory. This targetedprogram of metrics has increased management confi-dence in laboratory automation efforts, helped anticipatedata processing bottlenecks, and highlighted end-usersupport needs. These metrics have been automatedthemselves wherever possible.

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Strategic planning for bioanalytical automation:managing growth successfully

who understand our products and our requirements thatget systems on line.

Julie j. Tomlinson (Glaxo, Inc., Research Triangle Park, NC,t:SA)

Bioanalytical Automation rapidly expanded at GlaxoInc. through the first half of 1990. During that growthperiod it became apparent that to continue to besuccessful, the growth ofthe technology and the resourcesrequired it must be planned and managed. A strategicplan was prepared to determine where we are, where we"are going, and what we need to do to get there.

The plan describes the goals of the BioanalyticalAutomation Group and the most important requirementsfor achieving those planned goals:

(1) Long-term management commitment.

(2) Trained, dedicated personnel.

(3) Quality facilities.

(4) Teamwork.

(5) Investment in automation-compatible equipment.

The strategic plan was a year old at the time of thispresentation. The progress of the plan was discussed.

Managing robotics in the generic pharmaceuticalarena

Assembling an effective robotics organization requiresprework on the part of management. There must be aclear vision of the specific types of activities the group willperform. This vision can be used to establish a skillsprofile for the members ofthe team. The size ofthe team isalso important to assure momentum towards success.The author’s experience suggests at least four people arerequired to provide the variety of skills and keep thegroup going.

Each member’s personality type is an important compo-nent of establishing a new team. In robotics, one of themost critical talents is the ability to work on long-termprojects that constantly present new challenges. Thegroup members need to balance consistency of purposewith the ability to creatively solve a variety of problems.The group will not be effective in delivering newtechnologies unless they have the talent to train thenovice in a highly technical environment.

People who are successful in automation development areunique. They should have the ability to work comfortablyin a logic based environment, to become very creative ondemand, and to communicate highly technical infor-mation effectively. People do not usually possess all theseskills, providing their manager with challenging coachingopportunities.

Marianne Scheffler (Danbury Pharmacal, Inc., Carmel, NY,USA)

Danbury Pharmacal introduced robotics in 1987 as a wayin which to optimize throughput, reliability, and cost.Through the company’s experience with robotics, thispaper gave an overview of various issues, such asacceptance by upper management as well as on thechemist level, political confrontation, validation pro-cesses and product throughput.

Selection criteria for laboratory robotic applicationpersonnel

Peter W. Rulon (Advanced Technology and Validation Section,Quality Assurance Department, Norwich Eaton Pharmaceuticals,Inc., Norwich, NY 13815, USA.)Norwich Eaton Pharmaceutical recognized the benefits ofusing automation systems in the laboratory over sevenyears ago and created a robotic development area withinthe analytical method development group. Since thattime, they have placed into routine operation eightcomplete robotic systems and a large number of semi-automated systems.

This level ofactivity has provided many challenges for theautomation group. The success of this group has beenvery dependent on the number of talents of peopleworking these assignments. You can have the bestequipment and the vendor’s promises of success, but ourexperience has shown over and over again, it is the people

Helping laboratory automation live up to its promise

Gary W. Kramer (National Institute of Standards and Tech-nology, Chemical Science and Technology Laboratory, Gaithers-burg, MD, USA)The demand for chemical analyses is growing at anexponential rate. However, the advancement of labora-tory automation is progressing, at best, linearly. Why?We all want to be able to answer analytical questionsbetter, faster, cheaper, more safely, and with less personaldrudgery. Laboratory automation has long promised todeliver such benefits. But building automated systems isstill too difficult. Although some progress has been made,the goal of totally automated analyses and laboratoriesremains tantalizingly distant. How can we make thecreation of automated analyses simpler, less expensive,and less risky to undertake?

Standards can help. To interconnect chemical analysisequipment to form automated systems, we need tominimize the amount of custom development required.Standarizing the manner in which we control instrumen-tation and the ways in which we move data, samples, andmaterials through the system can greatly facilitate thefabrication of automated systems. Adherence to suchstandards will allow the creation of automated buildingblocks that can be easily interconnected to analyticalsystems.

Another impediment to the development of laboratoryautomation is the current shortage of people with the

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rhe 1991 International Symposium on Laboratory Automation and Robotics: Abstracts

requisite skills to specify, design, create, install, modify,customize, update, and maintain such instrumentation.Expertise in chemistry (or biochemistry), mechanicalengineering, electrical engineering, and computer scienceis required. Since it is rare to find individuals with thiscomplete skill set, larger organizations have set up groupsto handle automation development. Smaller operations,which cannot afford the luxury of such a multi-talentedgroup or find an automation Renaissance man, face astruggle in their automation efforts.

Vax mainframe computer. The selected colonies are thentransferred into a module containing fermentationmedium by a Puma 560 Robot (previously reported atISLR 1988). After incubation, the flex #2 workstationadds methanol to each vial and transfers aliquots of thebroth into a large number of microtitre plates. Theseplates are subsequently assayed for the presence of activecompounds. Additionally, the flex workstations cangenerate genomic libraries in E. coli, purify plasmid DNAand transform suitable biophores.

The Consortium on Automated Analytical LaboratorySystems (CAALS) was created at NIST to foster thedevelopment of laboratory automation. Its programs,including efforts to establish automation standards and toaugment the pool of automation practitioners, are beingsupported by leading users and manufacturers of labora-tory automation. By working together on problems thatcannot be solved by a single organization alone, we planto bring the benefits of automation into the laboratorymore quickly.

BIOLOGY/BIOTECHNOLOGY

The use of robotics in a cosmetic microbiologylaboratory

Steven Schnittger (Estee Lauder Research & Development,Melville, NY, USA)The preservative challenge test is used to determine theefficacy of the preservative system in a cosmetic formula-tion. This method is a highly labour intensive, repetitivetask, which utilizes many man hours. For this reason, arobot was developed to perform this function and free themicrobiologist to perform other functions.

The isolation of unique microorganisms from soilcommunities

Lynda Ford, Dan Bowden, Heather Boll, Walter Raas, ,JoeLykins, Jacob Zynger, Linda Bergin, Quensetta Lucas and OtisGodfrey (Lilly Research Laboratories, Eli Lilly & Company,Indianapolis, IN, USA)The screening of microbial colonies for the production ofnew pharmacologically active compounds is a labourintensive, integrated process consisting of colony isola-tion, fermentation, sample preparation and assay. In theflex 4/’ workstation a Mitsubishi Movemaster EX Robotweighs, sonicates, dilutes, and delivers individual soilsamples onto a series of different agar-containing plates.Following incubation and colony growth, the plates arescanned by the vision workstation. An ASCII filecontaining colony measurements and locations is pro-cessed by a fortran program which eliminates bacterialcontaminants using rules relating their size, % lighttransmission and uniformity of reflected light. Rules forpattern recognition are then employed to eliminatereplicate colonies from each soil sample. In addition,rules for the selection of rare colonies are generated afteranalysis of data files stored in an S 1032 database on the

Automated procedures for the determination ofenzymatic activity

Richard Harrison and Maria Izquierdo-Martin (Merck Sharp &Dohme Research Laboratories, P.O. Box 2000, RY-80N-A58,Rahway, NJ, USA)

Kinetic characterization of enzymatic activity has longbeen a labour intensive and time-consuming process. Afully automated robotic system was designed for thekinetic evaluation of enzymatic activity. The roboticsystem is a standard Zymate robot interfaced to anHPLC. Three unique features were incorporated into theapparatus; a low dead volume HPLC interface wasdesigned which minimizes the loss of precious, expensivereagents, a device which allows solutions of temperaturesensitive biological samples to remain accessible on thework surface at 4 C, and a temperature controlled rackwhich keeps reaction mixtures within 0.1 C of the settemperature.

With this robotic system, it was possible to determinekinetic constants for several enzymes. To illustrate theutility of the robotic system, kinetic evaluation of themetalloprotease Stromelysin was presented. Standarddeviations of initial velocities measurements are lowerthan when performed manually. The kinetic constantsKm and Vma are defined more precisely than with apreviously described manual method (Anal. Biochem.,18(I, 1989, 110), at one fifth the time. This is the firstreported fully automated chromatographic system cap-able of determining enzymatic kinetic parameters.

Automation for mapping of the human genome

Mary M. Blanchard, David Sloan and Volker Nowotny(Washington University School of Medicine, St. Louis, MO,USA,)

With the biochemical prerequisites at hand, the problemof establishing a map of the human genome that consistsofabout 3 billion basepairs was tackled. First, the humanDNA is kept in a library consisting of yeast clones,specifically, Yeast Artificial Chromosomes (YAC). EachYAC contains a piece of human DNA up to a lengthexceeding a million basepairs. This procedure allows forthe maintainance and propagation of the human DNA.Second, the Polymerase Chain Reaction (PCR) amplifiesshort tracts of specific DNA from within a very complexmixture of DNA sequences, making only this specificDNA visible. Thus, through the use of PCR, it is possibleto recover a clone from the library which contains the

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DNA sequence which is of interest to the investigator. Inorder to implement PCR in the screening of the library,the DNA of all the clones must be purified and arrayed ina pooling scheme. Then, one may identify positive clonesthrough a binary screen of this scheme.

Mapping of the human genome or human chromosomesthus becomes the repeated screen of the library fordefined sequences (Sequence Tagged Sites; STS). Screen-ing data will provide information about the order of theclones, thereby generating a chromosomal map. For asufficiently dense (on average one STS each 50 000basepairs) coverage of the genome, about 3000 STS’s perchromosome are necessary.

To automate the biological experiment (PCR), we utilizea robotic workstation that is based on the BIOMEK 1000system (Beckman Instruments, Palo Alto, CA 94304).Necessary modifications involve the increase of availabletip boxes to the robot’s envelope, the design and buildingof a thermocycler capable of handling 576 samples perround and a storage unit to have access to DNA samplesofabout 60 000 yeast clones. The major effort involves theassembly of software for the control of this machine toallow a completely automatic screen integrating thou-sands of pipetting steps and the transfer of hundreds ofmicro titre trays each day.

increased by a factor of three, labour decreased by halfand precision of results significantly improved. The samebasic laboratory robotic system is currently being used inother types of pesticide-soil research. For instance, thebatch equilibration method used to characterize pesticideadsorption-desorption was automated This methodinvolves equilibrating an aqueous pesticide solution withsoil for a period of time, separating the solution from thesoil, adding solution without pesticide to the soil andrepeating this process a number of times. After each step,the pesticide is extracted from each solution and ana-lysed. The experimental variables are soil type, pesticidesolution concentration, aqueous solution composition,and equilibration time. As part of a typical adsorption-desorption experiment, soil samples (three soil types, fourpesticide solution concentrations, three replicates) areprocessed after successive 3 h equilibrations over a 72 hperiod. The experimental procedures for the elucidationof binding mechanisms of pesticides to soil and theoptimization of parameters for extraction of pesticidesfrom soil have also been automated.

Zymark robotic automation of EPA Method 505:analysis of organohalide pesticides and commercialpolychlorinated biphenyls (PCB) products in waterby microextraction and gas chromatography

ENVIRONMENTAL

Laboratory robotics applications in environmentalresearch on the fate of pesticides in soil

William C. Koskinen, LeEtta J. Jarvis and Robert H. Dowdy(USDA Agricultural Research Service, and Soil ScienceDepartment, University of Minnesota, St. Paul, MN, USA)

Characterization of the environmental behaviour ofpesticides in soil entails field studies and laboratoryexperiments under controlled conditions. Field experi-ments to determine the dissipation and movement ofpesticides and laboratory experiments to determine theeffect of environmental factors such as soil temperatureand moisture on pesticide degradation often requirethousands of samples to be analysed. The analysis is amulti-step process including: extraction of the pesticidefrom soil, sample clean-up and preparation, identifica-tion, and quantification. The extraction ofpesticides fromsoil and subsequent sample preparation is the limitingstep in the analytical procedure. A robotically automatedprocedure has been developed by the authors for theextraction of a variety of herbicides from soil andsubsequent sample preparation. The procedure includes:adding extraction solvent to soil; mixing the soil-solventsuspension; centrifuging the suspension; removing super-natant solution containing pesticide from soil; evaporat-ing supernatant down to water; absorbing pesticideremaining in water onto solid sorbent; stripping pesticidefrom solid sorbent; placing final pesticide solution intovial; and capping the vial for GC or HPLC analysis. Withserial processing of samples by the robotic system, asopposed to manual batch processing sample throughput

Russell M. Spencer, Joseph A. Poland (CT Dept. of HealthServices Lab Bureau, Hartford, CT, USA.), and Marsha A. Paul(Zymark Corporation, Hopkinton, MA, USA)

Zymark robotics are uniquely qualified for environmentalwater testing through the efficient automation of EPAMethod 505. This method consists ofextracting 35 ml ofawater sample with 2 ml of hexane. Aqueous calibrationstandards are extracted and analysed in parallel with thesamples in order to compensate for extraction losses. Asingle injection of4 tl of the hexane extract is made into a

gas chromatograph with one injection port split toaccommodate two different capillary columns utilizingdual linearized electron capture detectors.

Zymark automation performs all the required laboratorysteps of the method completely and consistently fromsample acquisition to final analyte identification, confir-mation and quantitation. The Zymark PyTechnologysystem is composed of a concentric ring of specificfunctional modules serviced by a central Zymate IIrobotic arm. Laboratory processes including aspiration,weighing, mixing, extraction, transfer and injection intothe gas chromatograph have been integrated into a fullyautomated system under command of the System VController.

Certification and validation of the system was achievedby multiple analyses of proficiency sample matrix spikessupplied with EPA Water Pollution Study NumberWP023. Seven different pesticides including aldrin,dieldrin, pp-DDD, pp-DDE, pp-DDT, heptachlor, andheptachlor epoxide were analysed. Follow-up studies arein process for 14 additional pesticides and 7 PCBs. Linearregression curves plotted for aqueous multilevel extractedstandards (7 levels) yielded a correlation equal to or

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better than 0"9997 for the above pesticides over aconcentration range of 0"04 to 30 ppb in water. Finalconcentration results for eight replicate analyses of eachof these compounds all fell within EPA AcceptanceLimits. Mean per cent recovery was 92% or better of thetrue values for five pesticides, the exceptions beingheptachlor at 84% and dieldrin at 88%. Other statisticalconsiderations include a standard deviation range of0"02-0"08 and a variation from 42-11 for thesecompounds.

Significant savings in terms of laboratory resources arerealized when comparing the Zymark AutomationSystem with manual procedures. Automated analysis of40 water samples requires approximately 40 min to set upand run on the robotic system as compared to 240 min tocomplete manually. This is a 600% performanceefficiency rating for the Zymark system. In addition,automation provides for completion of the assigned taskduring normally non-productive hours in the eveningsand on weekends and holidays. In conclusion, reliability,sensitivity, precision and accuracy of pesticide analysisby EPA Method 505 combined with 600% greaterefficiency over manual extraction techniques are thehighlights ofthe Zymark Automation and PyTechnology.

Design and development of a robotics based aciddigestion system for environmental sample prep-aration: evaluation and characterization of aciddigestion procedures for graphite furnace AA andICP analysis

Iqbal Dawood Tabni, Gray Ward (Enseco Inc., QualityAssurance and Health & Safety Division, Somerset, NJ, USA),Debra Hasford, Erik Natkin and Maxine Pitard (Enseco-RockyMountain Analytical Laboratory, Denver, CO, USA)A Zymate II laboratory robot system was used tocompletely automate the acid digestion procedures formetal analysis. The Robotics Based Acid DigestionProcessor was utilized to prepare aqueous and soilsamples for Graphite Furnace AA and ICP analysis. Thesalient features of this application are complete automa-tion of acid digestion procedures for:

(1) Different matrices.

(2) Analysis type.

(3) Test type.

In this report the feasibility of a completely automatedacid digestion procedures based on a robotics system forroutine use was characterized and evaluated. A compari-son of analytical data obtained from the manual pro-cedures and automated system was presented.

Automated environmental extract clean-up with theBenchMate workstation

Richard A. Kern, Kevin O’Mara, Shirley Yaikow (MidwestAnalytical Services, Detroit, M148201, USA), Louis J. Cercone,Eric Andersen, Lisa Moore (EnviroTest Laboratories, Inc.,Newburgh, NY 12550) and Mark Cava (Zymark Corporation,Hopkinton, MA 01748, USA)

Traditional extract clean-up methods require manualdilutions, filtrations, injections, column conditioning,and reagent additions. This paper presented:

(1) The automated clean-up methods.

(2) The machine-generated audit trail of volumetricaccuracy and precision.

(3) The GC/MS results for the analytes of interest.

Two different applications of the BenchMate wereexamined: the traditional approved SX3 Bio Beads(EnviroTest); and the new high pressure column (Mid-west Analytical Services).

Environmental extracts were analysed for semivolatiles,pesticides, and PCBs in waste oils. The BenchMateWorkstation performed weighings, dilutions, filtration,and injection for GPC clean-up, florisil cartridge clean-up, and acid clean-up.

Fast turnaround, good data, and low cost, arejust a few ofthe benefits that have been realized for these analyses.

Robotic system for aqueous environmental samplepreparation

M.J. Zoellner, H. W. Emmel and L. D. Nelson (Dow ChemicalCompany, Midland, MI, USA)A robotic system for the automated preparation ofaqueous environmental samples has been successfullyimplemented in the author’s laboratory. The system isdesigned for preparation of aqueous sample types: riverand well waters, surface waters and effluents. The samplepreparations which are possible include digestion and/orconcentration that follows EPA-600/4-79-020 method-ology for trace elements via inductively Coupled Plasma(ICP); the determination of arsenic and selenium viahydride AA; and the determination of mercury via coldvapour AA.

Heating the sample either for digestion or volumereduction purposes has been traditionally done using hotplates requiring almost constant attention to prevent anyone sample from going to dryness which would volatilizeany metals present thus rendering the analysis useless. Itis this unpredictable non-uniform heating by hot platesthat makes it a poor choice for automated samplepreparation. The approach taken for this application wasto use a water bath for heating the sample. The waterbath provides for a uniform 98 C heat applied to all thesample tubers which minimizes sample bumping andreduces the chance of cross contamination. This arrange-ment also eliminates the need to constantly monitor theevaporation progress allowing for unattended use. Sincethe procedure is a timed event, the program controlallows for the added capability to use the robot arm forother purposes such as a serial dilution procedure whilethe samples reside in the water bath.

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A high temperature heating block (300 C) is used for theAs, Se preparation procedure to allow the samples to betaken down to a fuming acid condition. The tubes aretransferred to a fan cooled steel rack and allowed to coolto room temperature before returning to the final samplerack. This minimizes any accidental exposure to hot acid-fuming tubes since the robotic system is designed to

operate utilizing a standard laboratory fume hood.

Development of laboratory robotics at FDA: auto-mated system for the determination of natural toxinsand pesticides

Allen S. Carman, Jr., Shia S. Kuan (Natural Toxins ResearchCenter), Kenneth V. Miller and Humberto G. Guerrero (DistrictLaboratory, U.S. Food and Drug Administration, 4298 ElysianFields Avenue, New Orleans, Louisiana 70122, USA)Because of the reprogrammability of robot automatedtasks, this approach to automation has been calledflexible automation. This feature makes this type ofautomation particularly attractive to laboratories such asthose in the FDA requiring increased productivity, buthindered by a frequently changing and often unpredic-table, heterogeneous workload. To evaluate the potentialof flexible automation, the Natural Toxins ResearchCenter (NRTC) developed a workstation using a ZymateI System for the HPLC determination of solanaceousalkaloids in potatoes and the determination of aflatoxinsin milk and other matrices. This system was describedwith emphasis on a column liquid level sensor whosedevelopment was critical to the successful completion ofthe aflatoxin method and an HPLC injector developed in-house to save costs. Complete changeover of this systemfrom glycoalkaloid to aflatoxin assay can be accom-plished in less than 2 h.

A second workstation, using a PyTechnology System, isbeing used to automate a pesticide residue screeningprocedure developed by the District Laboratory specifi-cally for automation and the aflatoxin procedure de-scribed above. Comparison of recoveries of 15 pesticidesby this procedure with a well-established screeningprocedure shows that this procedure compares favour-ably with the established procedure. The screeningprocedure and the robotics system were described. Thedesign and construction in-house of a pneumatically-operated transfer arm, and the modification of a Zymarkevaporation station to provide controlled evaporationwere also presented.

Single lab evaluation of a robotic microwave diges-tion system for preparing aqueous samples byMethod 3015 and solid samples by Method 3051

Daniel C. Hillman, Piotr Nowinski (Lockheed Engineering andServices Company, 1050 East Flamingo Road, Las Vegas, NV89119, USA) and Larry C. Butler (Environmental Monitoring@stems Laboratory, U.S. Environmental Protection Agency, LaaVegas, NV 89114, USA)A robot system has been developed to perform microwavedigestions of environmental samples. The system iscomprised of a Zymark robot with several custom

PySections and a CEM microwave digestion oven.National Institute of Standards and Technology (NIST)supplied the software to interface the CEM ovenoperation with the Zymark robot operation. The robotmicrowave digestion system automates one of the mostlabour intensive steps in inorganic analysis. In a conven-tional manual digestion procedure, analyst attention andinput is required throughout the process, from weighingsamples, adding reagents, monitoring the digestionprogress, performing final dilutions, to transferringdigestate sample bottles or instrument autosampler vials.Critical throughout a manual procedure is accuratelyrecording the digestion data. With the robotic microwavedigestion system, operator input is limited to addingsample to digestion vessels and transferring digestate to

storage containers after digestion. All digestion steps(reagent addition, matrix spike addition, reagent blankpreparation, loading and unloading the microwave oven,and final dilutions) are performed by the system.Digestion protocols are downloaded from a data filecontaining the protocol data. By using barcode labels andbalances interfaced to the system, all digestion data isrecorded automatically in a text file. The operator is not

required to record any data.

Currently, two systems are being evaluated. One systemis located in Las Vegas, Nevada and the other inManchester, Washington. The primary differencebetween the systems is in the digestion vessel. The LasVegas system uses the single-wall Teflon digestion vesselwhile the Manchester system uses the newer double-wallTeflon-lined digestion vessel. The Manchester systemalso further automates the process by dispensing sample(after digestion and dilution) directly into vials held in anICPES autosampler rack.

This paper described the evaluation of both systems to

perform digestions of both aqueous and solid samplesfollowing SW846 Methods 3015 and 3051. Resultsincluded estimates of both within and between labprecision and bias. Also, general characteristics of thesystem were described (speed, reliability, data output).

Automation of an acid digestion process for watersamples

Gerald L. Hoffman (National Water Quality Laboratory, USGeological Survey, Arvada, CO, USA)

The present US Geological survey manual method ofsolubilizing trace metals associated with waterborneparticulates is a mild acid extraction procedure. Toautomate this procedure, it is necessary to change themanipulations and the conditions of the acid extraction.These changes are made not only to automate theprocedure, but also to lower the blank values for several ofthe metals determined in the acid digested water samples.It is important, however, for the automated procedure tostatistically duplicate the same acid-extraction efficien-cies for trace metals as the manual procedure, because ofthe large historical data base generated with the manualprocedure. Because of these requirements, the quantityand type of acid (2’5 ml of concentrated HC1 per 100 ml

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of sample) is maintained. Also, the use of Whatman* 42filter paper to filter the acid digested samples is continuedin the automated procedure. Major changes to themanual procedure are as follows:

(1) The acid is added directly to the sample bottle,instead of transferring the sample to a beaker andthen adding the acid.

(2) The closed sample bottle is heated in an oven for 8 hat 65 C, instead of heating the beaker on a hot plateat 90 C for 2 h.

Automation in pesticide residue analysis by a robotsystem

R. Brennecke (Bayer A G, Business Group Plant Protection,Institutefor Product Information and Residue Analysis, Monheim,Bldg. 6610, D 5090 Leverkusen, Bayerwerk, Germany)

Generation ofcrop residue and environmental fate data isan important part of both the development of a newpesticide and the support of existing products. Thedevelopment process spans a number of years andgenerates a large number of different samples, from cropsto soil and animal tissues, all requiring analyses down tolow levels, typically from 0"01 to 0’05 mg/kg. Thedemand for this type of trace level analysis has increasedstrongly over the last five years at the author’s Institute.

For handling all the samples it was decided to automatethe clean-up steps in residue analysis. More uniform andsimplified residue methods have been developed first,because the existing methods varied greatly and were toocomplex for automation. A liquid-solid extraction ondiatomaceous earth followed by an adsorption chroma-tography on silica gel was the resulting clean-up pro-cedure, which has been proven to be useful in routineanalysis with a large number of plant materials andbeverages and also for different pesticides. The next stepwas the adaptation of this procedure to a Zymate IILaboratory Automation System. A description of theadaptation and the validation of this clean-up procedureby a robot system was given.

CHROMATOGRAPHY SAMPLE PREPARATION

A validated liquid chromatographic assay for CI-988in human plasma with batch robotic samplepreparation

E. L. Johnson, K. Rutkowski, J. P. Hinton and D S. Wright(Parke-Davis Pharmaceutical Research Division, Warner-Lam-bert Company, Ann Arbor, MI, USA)CI-988 is cholecystokinin-B receptor antagonist which iscurrently under investigation as a potential antianxietyagent. Human studies will involve analysis of CI-988levels in a few hundred to possibly several thousand

The use of brand names in this abstract is for identification purposesonly and does not constitute endorsement by the US Geological Survey.

samples. Batch robotic sample preparation will be used toincrease total sample throughput as well as decreaseanalyst time.

The method involves batch robotic sample preparationwith a Zymate Laboratory Automation system and acustom vacuum box to hold the solid phase extractioncartridges. Up to 144 individual cartridges can beprepared in a batch mode. Solvents are added to thecartridges via a liquid distribution hand using either amulti-valve FMI pump or the master lab module as thesolvent source. Vacuum, controlled by the power andevent controller, is used to aspirate solvents through thecartridges. A TurboVap LV was used to evaporate eluentfrom the cartridges prior to analysis by HPLC withfluorometric detection.

The method has been validated and is suitable for routineanalysis of CI-988 in human plasma over the concen-tration range of0"25 and 500 ng/ml. Sixty samples can beextracted in 55 min. The rapid rate of sample extractionallows a single robot to be used by more than oneindividual during a routine work day.

Analytical chemistry in the toxicology laboratory: aversatile robotic sample preparation system

R. M. Kannuck, D. A. Redder and P. T. Hardesty (E.I. DuPontDe Nemours and Company, Haskell Laboratory for Toxicologyand Industrial Medicine, Newark, DE, USA)

Toxicology testing under EPA and FDA guidelines mustbe conducted according to published Good LaboratoryPractices (GLPs). These guidelines generally mandatethe verification of administered dose levels as well asconfirmation ofhomogeneity and stability of test materialin the dosing vehicle. The duration of toxicology testsmay range from two days to two years, with samplesbeing submitted in batches for analysis at varyingintervals throughout each study. The variety of testingrequires analysis for the same compound in completelydifferent matrices (animal chow, methyl cellulose,organic solvent) over concentrations ranging from severalper cent to parts per billion.

Such a dynamic environment does not seem at first glanceto be a likely candidate to benefit from robotic automa-tion, since there is no single assay or method that can beoptimized once and then used respectively for largenumbers of samples over a long period of time. However,because the total number of samples from all concurrentstudies is large and there are many repetitive operationsinvolved in the multiple methods that are supported, thissort of laboratory can still be amenable to the productiveuse of robotics.

Haskell Laboratory has developed a flexible system basedon a Zymark PyTechnology System that can be re-configured easily to implement methods that change on a

daily basis. The system can perform the subsampling,extraction, filtration and dilution functions necessary

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prior to chromatographic analysis. Additionally, systemevaluation routines and documentation necessary tocomply with GLP guidelines has been incorporated.

overnight operation allows the user to maximize produc-tivity. Precision and accuracy of the robotic proceduresare comparable to those obtained with manual methods.

Comparison of workstations for liquid-liquidextractions

j. K. Bolon and R. F. Arrendale (Drug Metabolism Department,Glaxo Research Institute, Research Triangle Park, NC, USA)Automated sample preparation was considered as ameans of enhancing productivity. Three automatedworkstations (the Tecan 5000, the Waters Millilab andthe Zymark BenchMate) were evaluated for use in aliquid-liquid extraction procedure to quantify G187084{methyl-3- [4-methoxycarbonyl-4- 1-oxopropyl)phenyl-amino]-[piperidine]propanoate}, an ultra-short actingopioid, from whole human blood. Levels of the opioidwere determined by capillary GC/MS/SIM. Accuracywas determined by weighted linear regression analysis ofthe calibrated standards. The BenchMate, Millilab andTecan 5000 had correlation coefficients of 0"9991, 0"9981,09978, respectively. Precision was measured by thecoefficient of variation (r) of the interpolated standardcurve concentrations. The coefficient of variance at theng/ml level for the BenchMate, Millilab and Tecan

5000 were 10’7%, 8"3%, 50%, respectively.

Medicated feed assay applications using a fullyautomated PyTechnology robotics system

L. C. Erhart, L.J. Kostek, J. E. Curley, A. Grizzuti and M. P.Synder (Pfizer, Inc., aroton, CT, USA)

The Analytical Research and Development Departmentof Pfizer Central Research has provided support totoxicology research by assaying medicated feeds to assureappropriate drug content during dosing. Historically,feed assay methods have been laborious, time-consumingand difficult when drugs are extracted from complex feedmatrices.

To improve laboratory efficiency, several automatedprocedures have been developed for the analysis of drugsin rodent feeds. The robotic system performs all thenecessary steps to determine the concentration of drug inanimal feed. The robot tares sample containers to whichan operator adds medicated feed samples. After deter-mining the sample weight, the robotic sample schedulermanages the robot’s time and motion according to pre-setparameters to efficiently handle up to eight samples at atime. Extraction solvent is added and the containers areshaken to extract the drug. Serial dilutions are made intoindividual tubes. The drug is separated from the feedmatrix by liquid-solid extraction or fatty oil precipitation.The solution is filtered and injected onto an isocraticHPLC system.

Application of these procedures with drugs in medicatedfeeds is discussed. Although the robotic method isinherently slower than the manual method, unattended

Automated sample preparation of whole blood forcyclosporin using the BenchMate

William Holman and Robin A. Felder (The University ofVirginia Health Sciences Center, Department of Pathology,Charlottesville, VA, USA)

Cyclosporin A (CsA) is currently the most selectiveimmunosuppressant used clinically for the prevention ofgraft versus host disease following organ transplantation.The narrow difference between renal toxicity of anexcessive dose and inadequate immunosuppression by alow dose makes constant monitoring of patient bloodconcentrations a necessity. Since immunosuppressionrequires continuous presence of CsA, organ transplantrecipients usually continue to take CsA for the rest oftheirlives.

High performance liquid chromatography (HPLC) is themethod of choice for measuring CsA. However, HPLC isplagued by low throughput since many manual prepara-tive steps must be performed prior to analysis. Alter-native methods, which include several variations ofmanually performed or automated immunoassay haverelatively high imprecision, interferences from metabolicproducts and tremendously high cost. Therefore, auto-mation ofCsA analysis by HPLC should result in an easyto perform cost effective method.

The Zymark BenchMate was used to automate much ofthe sample preparation involved in solid phase extractionof CsA. Whole blood specimens were collected fromvenipuncture into heparinized glass tubes. Samples wereplaced on the BenchMate in the specimen holding areaand a C 18 Bond Elut column was placed in the top of thesample tube. Specimen extraction from this point washandled automatically by the BenchMate. The samplewas injected manually into the HPLC system forquantification.

Similar recovery of CsA was found when manual andBenchMate methods were compared for three levels ofcalibrator. Analytical imprecision was smaller for theBenchMate when compared to manual extraction.Within run precision was found to have a coefficient ofvariation (C.V.) of less than 8% at either concentration.Between run precision was determined by performing aBenchMate extraction on two specimens each day for 20days. Between run precision was less than 9% C.V.Eighteen patient specimens were extracted eithermanually or using the BenchMate and the resultscompared by linear regression. There was good agree-ment between the manual and BenchMate extractedspecimens.

The BenchMate is useful for reducing the manual stepsnecessary to perform HPLC analysis of CsA. TheBenchMate has a throughput of 10 min per sample whichwould allow up to 48 samples to be extracted in an 8 h

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shift. Since extractions are performed serially by theBenchMate, samples can be added to the BenchMate asthey are received in the laboratory obviating the need forcut-off times for analysis.

A generic robotic sample processor for HPLCanalysis

L.j. Lorenz, E. C. Jensen, M. E. Hinshaw, D. A. Jackson andJ. Zynger (Lilly Research Laboratories, Eli Lilly and Company,Lilly Corporate Center, Indianapolis, IN, USA)Laboratory robotics generally follow a course where arobotic system is designed to mimic a previously devel-oped manual procedure. This presents a problem sincemost procedures vary. Therefore, a different system isneeded for each application changes or ceases to exist.

An alternative approach for utilization of laboratoryrobotics requires the development of a general purposerobotic system. Applications are then developed to fit therobotic system. Such an approach allows for roboticsystems which can handle a large volume number ofdifferent methods without alteration of the roboticsystem.

Such a system has been developed as a sample processorfor HPLC. Today, many samples which have very limitedsolution stability are examined by HPLC. This poorsolution stability limits the ability to use classicalautomated autoinjectors for servicing an HPLC instru-ment. A robotic system can overcome this problem bypreparing the samples just prior to analysis.

The robotic system is designed to make use of commer-cially available devices where possible. The system willhandle liquid samples when desired but is primarilydesigned to handle dry powders, capsules, or tablets. Thesystem will process a dry sample which is presentedpreweighed in a test tube. A solvent is added and thematerial is put in solution through a series of shaking andsonication steps. The sample can undergo a subsequentdilution step with mixing if desired. The solution is thentransferred to a syringe filter which filters the sample andalso acts as the driving force to push the sample throughselection valves to direct the sample to a specificinstrument and finally through a sampling valve to injectthe sample into the HPLC system. The robotic systemincorporates appropriate relays and logic to start anHPLC gradient to autozero detectors, and to start datacollection for the sample. The robot documents all keysteps in the process and documents all error recoveryattempts when a problem is detected.

GENERAL

Trials, tribulations, and triumphs in PyTechnology

Bruce Kropscott, Jeanne Hugo (Dow Chemical Company,Midland, MI, USA), and Shoreh Shabrang (Manteq Inter-national Corporation, Midland, MI, USA)The Zymate Laboratory Automation system has gonethrough considerable evolution since its introduction atthe 1982 Pittsburgh Conference. The first Zymate

robotics systems were able to perform a limited number oflaboratory unit operations (LUOs) such as weighing,pipetting, mixing and container manipulations. TheZymate I robots were easy to direct through a series ofpositions but each position on the table had to bemanually taught. Each LUO involved moving throughmultiple positions an.d actual applications used hundredsof positions, thus applications were physically labour-intensive to set-up and took weeks even months to fullyimplement. The custom CPU controllers were limited inthe amount of memory; had no inherent error detection;and LUO verification was left up to the individualprogrammer.

Since the first introduction, the robotics systems havebeen transformed through rapid improvements such asincreased speed, tactile and axis sensing and new LUOscapabilities. In the 1990s, PyTechnology RoboticsSystems have many pre-positioned peripheral moduleswith pre-programmed LUOs thus allowing a faster start-up time for standard applications. PyTechnology pro-gramming has extensive error checking to help the user toanticipate probable errors and has pre-determined rou-tines to recover from those common errors. Unfortu-nately, many ofour applications require custom modules;use more than the allotted number of available PySec-tions; use a variety of containers; and incorporate non-standard routines for the pre-programmed LUOsmodules.

This paper described (1) the advantages and constraintsof PyTechnology for custom applications; (2) efficient useof the working space by stacking racks and combiningLUO modules; and (3) basic philosophies andapproaches for custom software in the authors’ methoddevelopment system.

Automating test methods for evaluating chemicalwarfare protective materials

Raymond E. Andreotti (U.S Army Research, Development andEngineering Center, Soldier Science Directorate, Natick, MA,USA,)

The US Army’s research programme on the developmentof chemical protective clothing involves the evaluation offibres, foams, films and powders for sorptive and/orreactive activity toward chemical warfare agents. Experi-mental samples are subjected to droplets or vapours ofchemical agent simulants in closed containers for specifictime periods. The sorptive and/or reactive characteristicsof the sample are measured as a function of time byanalysing for the remaining simulant using a gaschromatograph of a UV/visible spectrophotometer.Liquid-state tests are also performed utilizing a Fluoride-sensitive electrode to detect the hydrolysis product of thereaction. Due to the variety and complexity of the tests, aswell as the toxicity of chemical agent simulants used,automation is a real challenge. This paper discussed howthe flexibility of three laboratory robots was utilized indeveloping automated test methods that improved safety,accuracy, and productivity under hazardous testingenvironments.

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Pouring difficult solids under diverse conditions

Stephen R. Metzner (Monsanto Agricultural Company, 700Chesterfield Village Parkway, St. Louis MO 63198, USA)

When working with solid materials, it is often necessaryto pour all or part of the solid from one container toanother. Often the weight of the solid is desired beforedissolution, extraction or subsequent handling. Thenature of solids may vary widely and contribute to thedifficulty encountered when a robot pours the material.

The need to keep containers capped may control whichcontainers are used to hold the solid. The nature of thereceiving container may also be determined by subse-quent analytical procedures.

Beginning with a powder pouring routine developed byZymark, this poster presentation described the successesand failures of robotic pouring of difficult solid materials.

Designs often require modifications of the hardware and/or modifications of the task. While compromises are oftenessential, they represent a risk for eventual dissatisfac-tion, and potential failure. A common compromise isgiving up space in a crowded laboratory. Newer designsare becoming more compact and vertically integrated.

Custom automation techniques

Ron Hunt (Helene Curtis, Inc., Chicago, IL, USA)Bench-top automation to enhance productivity in thelaboratory is, for the most part, available off the shelf. Insome cases, however, parts of the application, or even allof the application require custom designed workstations.This presentation will discuss the advantages anddisadvantages of in-house design versus contract vendordesign. Examples were given on the handling of heavywet.samples by use ofcustom fingers, the implementationof barcode readers for decision and alignment purposes,and the mechanics/control of custom workstations.

Polymer ashing- a new look at an old technique

Joe Cross, Ron Jones, Connie Lowe and Mitch Meyer (PhillipsPetroleum, Bartlesville, OK 74004, USA)

Ashing has long been a conventional technique forpreparing polymers for analysis of catalyst residues andtrace metal contaminates. It is a labour intensive, messytechnique, used to prepare samples for both spectroscopicand gravimetric procedures. The conventional techniqueinvolves 30 to 60 min burning in a crucible over an openflame, followed by a high temperature ashing of theresidue in a muffle furnace to remove all traces of carbonresidue. The open flame burn-off requires operatorattention and is a process that would be difficult toadequately automate. This report detailed the results ofan investigation of an alternate technique for ashingpolymers; direct ashing of the polymer in a hightemperature muffle, in order to arrive at an acceptableprocedure that eliminates open flame burn-off andsimplifies automation.

Designing automation for laboratories

Michael A. Roos (Laboratory Robotics Company, St. Louis, MO,USA,)

Over the years of designing laboratory automationseveral lessons have been learned from systems that weresuccessful, and equally important, from systems thatfailed to meet expectations. The automation designersthat are most successful today have learned these lessonsand are incorporating them into their system designs.

Several examples of design criteria and strategies werepresented. The most important aspect ofany design is thecareful and realistic planning of the specifications andexpectations. The next design step is the proper specifica-tion and selection of automation, and hardware. Systemdesign is shifting away from performing detailed tasks bythe robot, toward parallel operations that use the robot asa facilitator between sophisticated workstations.

PLENARY

Superior productivity- in the laboratory and beyond

Francis H. Zenie (Zymark Corporation, Hopkinton, MA 01748,USA)Today, the need for more productive laboratories is morecompelling than ever before. As we enter the 1990s,intense world-wide competition and increasing regula-tion challenge our laboratories to ensure: more preciseresults, unquestioned documentation, faster turnaroundand better utilization of skilled people.

Superior productivity demands the greatest economicvalues from our resources; people, time and capital. Notonly must modern laboratories improve their internalproductivity, they must enhance productivity throughoutthe organization.

Many leading businesses have embraced laboratoryautomation as a strategic foundation. They invested to

gain knowledge of new technologies and experienceapplying them- leading to impressive results. Thispresentation described specific case histories and drewpowerful conclusions from the common elements of thesereal-world examples.

POSTERS

System data management for automated tablet assayworkstations

Jeff Roberts and Michael Shirley (Bristol-Myers Squibb,Products Division, 9707 Chapel Hill Road, Morrisville, NC27560, USA)The authors’ purpose was to design a data managementsystem that would allow us to generate batch datasummary reports, as well as to capture critical system

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data, in order to continually monitor a CustomizedAutomated Tablet Assay Workstation via SPC Charting.

The critical data, from both sample preparation andsubsequent HPLC Analysis, is captured real-time by theSystem V Controller and a PE Nelson Interface. The datais then imported into Lotus 1-2-3, and is manipulated bya series of Lotus Macros.

The poster showed sample programs written in EasyLab,PE Nelson Chromatography, and Lotus Macros softwarelanguages. In addition, sample data and graphs werepresented.

Automated extraction of XDE-436 from dog chowreplaces soxhlet extractions

Cynthia N. Peck (Analytical Chemistry Laboratory, 1803Building, The Dow Chemical Company, Midland, MI 48674,USA).

XDE-436

An automated procedure for the extraction of XDE-436,an experimental miticide, from dog chow has beendeveloped using a PyTechnology robotics system. Thisprocedure reduces the analyst time required for theextraction procedure from 6 to 11/2 h for 40 samples andeliminates the use oftedious soxhlet extractions to removethe test material from the feed.

XCE-436 (4- (2- (4- (1,1-dimethylethyl) phenyl) ethoxy)-quinazoline) is currently undergoing toxicity testing indogs and rodents. Previously developed analytical meth-ods determined that XDE-436 is stable in rodent andavian diets for up to 30 days.

XDE-436 could be quantitatively extracted from spikedsamples immediately after mixing using a simple solventextraction. However, the recovery dropped below 90%about 12 hours after mixing. The feed samples oftenarrived for analysis late in the day, preventing same daymanual extraction without overtime work.

An automated method for solvent extraction of the feed,followed by centrifugation and dilution of the extractswas developed using a fully equipped PyTechnologySystem. The analyst can prepare the system for un-attended overnight extraction in less than h after thesamples arrive, allowing the test material to be removedfrom the feed within 12 h.

A highly automated fermentation extraction system

C. K. Marschke, L. R. Carter and D. D. Gleason (The UpjohnCompany, Kalamazoo, MI 49007, USA)The Upjohn Company’s Chemical and Biological Screen-ing unit was organized to collaborate with the company’sdiscovery units in exploiting natural products as sourcesfor new therapeutic activities. The system described hereprovides for the processing of large number of microbialfermentation broths and extracts.

A major need on one project was to extract eachfermentation broth with an organic solvent. The systemto do this was designed in collaboration with ourLaboratory Automation Support group. The micro-organisms were grown in specially designed, 45-wellcassettes and a Zymark laboratory robot performed theextractions. This system has many unique aspects butmost noteworthy is a multifunctional ’hand’.

The first step in the extraction procedure is the sequentialaddition of acetone and chloroform to each well of thecassette. For this operation, the hand is rotated to activatea paddle that is lowered into a well to stir thefermentation broth during addition of organic solvents.The robot then aspirates a several millilitre sample of theimmiscible organic solvent and places it in a conicalvessel. For each sample-transfer, a pipette manipulatoron the hand picks up a pipette tip, uses it, and thendiscards it. The holder for the pipette tips is also an airmanifold that directs heated air into the cups to evaporatethe solvents. With the solvents evaporated, the hand isrotated to use the aerosolizing tip for the addition ofreconstituting solvent. These reconstituted extracts aredistributed to various bioassay laboratories.

Integrated use of robotic systems in a high through-put approach towards drug discovery

Mark Beggs, John Major, Colin Bath and Tina Hayden (HighThroughput Screening Laboratory, Bioscience II, ICI Pharma-ceuticals, Mereside, Alderley Park, Macclesfield, CheshireSKIO 4TG, UK)A range ofrobotic systems are being used to identify novelchemical leads against selected targets against whichthere are no obvious starting points for medicinalchemistry programs based programs.

Randomly submitted samples from the company’s com-pound collection are solubilized in dimethyl sulfoxideusing a Zymate II System. The use of appropriatebarcoding technology permits the robot to solubilizesamples ofvarying weights to give stock solutions of fixedconcentrations. Working strength solutions are thenautomatically prepared from the stock solutions.

Assays are subsequently performed using a TecanRSP5072 robot utilizing Amersham’s Scintillation Prox-imity Assay Technology. Text compounds, controls andreagents are added to 6 x 16 well T-Trays which aresealed and counted in an LKB Pharmacia 1205 Betaplatescintillation counter. All data analysis is automatic and

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results are automatically uploaded onto a mainframerelational database.

Automation ofthese processes has resulted in a seven-foldincrease in assay throughput as compared to manuallyoperated screens. This throughput has been achievedwithout any further increase in staff.

An automated method for the determination of anew antiepileptic drug candidate (CGP-33,101) inhuman plasma using high-performance liquidchromatography

Linda A. Brunner (Development Department, PharmaceuticalsDivision, Ciba-Geigy Corporation, Ardsley, NY, USA)

An automated method utilizing laboratory robotics hasbeen developed and validated for quantifying concen-trations of a new antiepileptic drug candidate (CGP-33,101) in human plasma. The robotic system aliquotsthe biological sample, adds the internal standard (CGP-23,901), extracts the compounds from the basifiedbiological matrix (pH 12) into an organic phase(dichloromethane:methyl t-butyl ether) and concen-trates the extracts for reversed-phase, high performanceliquid chromatography (HPLC) analysis. The laboratoryrobot is directly interfaced to the HPLC system, and thedata are automatically collected and results calculated.Separation is achieved on a 3-um, Hypersil ODS (4"6 x50 mm) column with ultraviolet (UV) detection of thedrug and internal standard at 230 nm. Recovery andreproducibility assessments indicate good accuracy(overall mean relative recovery of 102’7%) and precision(coefficient of variation of 4.4 to 7"7%) over the CGP-33,101 concentration range of 50 to 4000 ng/ml, with a

quantification limit of 50 ng/ml. The method has beensuccessfully applied to a pharmacokinetic study in whichnormal volunteers received single, oral doses of 400-1200 mg CGP-33,101.

An automated sample preparation and radiochemi-cal HPLC analysis of Technetium 99mTc-teboroxime

R. Fisco, J. W. Sulner, D. Silowka, J. Troskosky, J. P. Zoddaand M.N. Eakins (Bristol-Myers Squibb PharmaceuticalResearch Institute Diagnostics Evaluation Department, NewBrunswick, N,] 08903, USA)

The sample preparation and radiochemical HPLC analy-sis of a new myocardial perfusion agent, 99mTc-Tebor-oxime (CardioTec), has been automated using a shieldedZymark V Robotic System. Product specificationsrequire 99mTc-Teboroxime to be reconstituted with mlof diluted eluate containing radiochemical concentrationof 100mCi 99mTc-pertechnetate (TcO4). The robotweighs a reaction vial, calculates the ratio of radioactiveeluate and 0’9% saline contain 100mCi 99mTcO4 in ml,delivers the volume into the vial, and constantly adjuststhe volume-ratio of eluate-to-saline for subsequent vialsby incorporating the decay factor for 99mTechnetium(T1/2 6 h) into each calculation. Following reconstitu-tion, the robot weighs each reconstituted reaction vial toconfirm the volume, vortexes the vials, heats the vials for

15 min at 100 C in a specially constructed lead shieldedblock, cools them to room temperature and then sequen-tially delivers a 0’2 gl sample into each of two HPLCsystems to determine the radiochemical purity of thesample. The reconstituted reaction vials are stored in alead block for 6 h and then reassayed by sequentiallydelivering a 0"5 gl sample into each HPLC system.

The advantages ofautomating this procedure include: (1)decreasing the amount of radiation exposure to labora-tory personnel; (2) relieving skilled staff from performingroutine, tedious tasks; (3) increasing productivity byallowing unattended operation for extended periods oftime; and (4) increased assay reproducibility by eliminat-ing human error.

Robotic sample preparation and HPLC analysis ofVerlukast in human plasma

Charles Lin,John Yeh-Kang Hsieh, Bogdan K. Matuszewski andMichael R. Dobrins.ka (Merck Sharp & Dohme ResearchLaboratories, Department ofDrug Metabolism, West Point, PA19486, USA)An automated sample preparation procedure was de-scribed for analysing plasma samples originating frommetabolic disposition and bioavailability studies. AZymatc I Laboratory System was modified and upgradedto prepare plasma samples for the quantification ofverlukast, a potent LTD4 receptor antagonist.

The robot performed all necessary steps for the plasmaprotein precipitation and then transferred the super-natants to the Perkin-Elmer HPLC system by directinjection. Verlukast and internal standard were separ-ated on a DyChrom Chemcosorb ODS-UH column (5 u,4"6 mm x 150 mm) using a mobile phase consisting ofmethanol and ammonium phosphate buffer. Detectionand quantification were carried out using a McPhersonFL-750 Fluorometer and an HP 3357 Laboratory Auto-mation System, respectively.

The system has been developed for unattended 24 hoperation in the dark due to photosensitivity ofverlukast.The overall accuracy and precision attained is 98’9 +0"2% for concentrations of 0’ to 5 g/ml. The analyticalmethod has been demonstrated to have the sensitivityand specificity necessary to quantify plasma concent-rations of the above compound following oral and I.V.administration of verlukast to normal subjects.

Laboratory automation study

Allan L. Greenberg (Laboratory Robotics Interest Group, NJ,USA) and Linda A. Brunner (New York-New England LabAutomation Interest Group, USA)Midwest Research Institute has conducted a survey onlaboratory automation and its implementation. Thesurvey was supported by both the Laboratory RoboticsInterest Group of New Jersey and the New York-NewEngland Lab Automation Interest Group.The results from more than 2000 responses were pre-sented. The information should provide users of labora-tory automation technology with some valuable infor-mation and insights.

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Development of a customized Zymate II system toautomate primary structure determination of recom-binant proteins

M. Reino, P. Diegelman, S. Pocchiari, M.J. Ehrke andJ. W.Cowens (Grace Cancer Drug Center, Roswell Park CancerInstitute, Buffalo, NY 14263, USA)Quality control procedures for insuring the identity andpurity of materials used as therapeutic agents arerequired by the FDA. The utilization of recombinantproteins in therapeutic applications is an area of rapiddevelopment; however, most of the procedures for thequality control ofsuch materials require specially trainedpersonnel and have not been automated. A ZymarkLaboratory Automation System is being developed toautomate the procedures used to determine the primarystructure of proteins and polypeptides, i.e. amino acidanalysis, enzymatic mapping, and sequencing. Toaccomplish this task, the automated system must success-fully carry out a series of discrete chemical reactions foreach analysis and since the size of sample available foranalysis may be small, the PICOTAG system developedby Waters Chromatography Division has been used asthe model for carrying out a series of chemical reactionson a microscale. In the PICOTAG system all reactionsoccur in a 6 x 50 mm Pyrex tube contained with areaction chamber under inert atmosphere; reagents areremoved after each reaction by a vacuum system. Toadapt this approach to the Zymate II System, ourlaboratory collaborated with Zymark to modify standardstations to accommodate the chamber-tube assembly, todevelop custom stations for vacuum evaporation andpurging with inert gases, and to write the new softwarerequired for the robot to interact with the modified andcustomized stations. It has been demonstrated that therobotic system can cari’y out each step required for theamino acid analysis procedure. A synthetic 10-merpeptide has been analysed on the customized Zymatesystem and the results are completely comparable tothose of the PICOTAG system. Studies are under way todetermine the sensitivity and reproducibility of theanalysis on the robotic system. Since the robotic system iscapable of performing a series of modular operations(pipetting, weighing, mixing, heating, evaporating, etc.)in any order, the automation of enzymatic mapping andprimary sequence determination are being developed onthe same system.

A robotic system for the automated DNA sequencingreactions

S. Kajie and Y. Mizuno (Torten Corporation, Saitama, Japan)The manual method for the DNA sequencing prior to theanalysis by automated DNA sequencer is a dull and time-consuming procedure, and the process needs to beperformed under dim environment for the light protec-tion offluorescent-!abelled dye primer. Completion of thesequencing reactions is one of the rate-limiting steps inautomated DNA sequencing.

To increase efficiency and reliability in automated DNAsequencing, the fluorescent-labelled DNA sequencingreactions have been automated including annealing

reaction, elongation/termination reaction, and sampleconcentration by ethanol precipitation. The instrument isa Zymate II Laboratory Automation System modified to,include handling of screw-capped microcentrifuge tube,high-speed refrigerated centrifuge, liquid sample process-ing, and thermal treatment. Reactions were performed inmicrotubes with screw cap, which prevented loss ofliquidby evaporation and ensured mixing of micro!itre quanti-ties of solutions by means of Vortex and brief centri-fugation. Sequencing samples were prepared by Sangerdideoxy method using E. coli DNA po!ymerase I Klenowfragment. The quality and reproducibility of the reac-tions were examined by an automated DNA sequencer,and sequence data of comparable quality to those ofmanual sequencing reaction was obtained.

Use of robotics for the automation of the angiotensinII receptor binding assay

Mary Jo Wilde,y, John Nygaard, Deborah Steinberg, MichaelGreenstein and William M. Maiese (Lederle Laboratories,Middletown Road, Pearl River, NY 10965, USA)In the pharmaceutical industry, high-throughput screen-ing is a key step in the discovery of novel agents withtherapeutic utility. Many detection systems, such asreceptor binding assays, involve repetitive procedureswhich can be readily conducted by laboratory robotics toachieve the desired throughput goals. An automatedprotocol for the [125](sar1, ilea)Angiotensin II receptorbinding assay, developed by Lederle’s CardiovascularResearch Section, is currently being run. A BeckmanBiomek SL robot is employed to accurately pipette testsamples, ligand, and receptor to the wells of a microtitreplate. After a timed incubation, all 96 wells of the assayplate are simultaneously harvested onto a filtermat andwashed by a Tomtec 96 well harvester. Receptor-boundligand retained on the filtermats is subsequently quanti-tated by scintillation counting on a LKB Betaplatereader. Data is automatically transferred to a database onLederle’s VAX mainframe for subsequent analysis,permanent storage, and report generation. For allautomation projects, criteria for acceptability, whichrequire the robot to meet or exceed the statistical limitsfor variability established for the manual version of eachassay, have been devised. In an initial assessment of thissystem, a random set of synthetic compounds wereevaluated at the rate of 350 samples per hour. Activecandidates were further analysed robotically to determinetheir ICs0’s. Assays relevant to all therapeutic areaswithin Cyanamid are under consideration for automationand subsequent testing of synthetic compounds andnatural products.

Robotics and automation of ligand binding assays

Randy A. Turner and Robin A. Felder (University of Virginia,Charlottesville, VA, USA)Ligand binding has become an indispensable tool in basicresearch laboratories and the pharmaceutical industry.Assays measuring the binding of fluorescent or radioiso-tope labelled ligands to cell surface receptors allowsensitive and rapid screening of drug potency andspecificity. Robots and the peripheral devices with which

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they interact can be used to automate various aspects ofreceptor assays. Dopamine receptors were studied in ratbrain and kidney using a variety of dopamine agonistsand antagonists in competition with 125I-labelled Sch23982. Identical assays were performed manually andwith a Hamilton Microlab AT pipetting robot. TheMicrolab AT was allowed to make serial drug dilutions,including tip wiping, tip changing, and sample mixing. Itwas also allowed to dispense samples and reagents ontomicrotitre plates. All other steps in the assays wereperformed manually. For a twelve drug assay, the robotwas more than six times faster diluting drugs than ahuman technician, and was more than eight times fasterthan a human in dispensing 50 ul aliquots of drug, tissue,competing ligand, and buffer into each well of a standard96-well microtitre plate. The competition curves gener-ated by both manual and machine assays were in goodagreement; however, the repetition standard deviationswere significantly lower for the Microlab AT (manual513 + 84 CPM, machine 485 + 20 CPM, p -< 0"05, N3). Use of the robot allowed smoother curves to begenerated with fewer repetitions (N 2 vs. N -> 4) andsmaller volumes (15 ul vs. 50 ul). Scatchard plots fromthe robot data yielded dissociation constants and maxi-mum receptor densities in agreement with literaturevalues. Precision and accuracy tests yielded coefficients ofvariation of 9% to less than 0’5% for 5 microlitrevolumes, depending upon the pipetting protocol; andsignificantly better values were obtained for largervolumes. Other aspects of radioligand binding automa-

tion, including the use of cell harvesters and microtitrebased scintillation and radioactivity counters, were alsodiscussed.

Handheld computer RS232C interfacing to an ana-lytical balance

Albert Marcus Morrishow and Wah Sang Lee (Mount SinaiSchool of Medicine, New Yok, NY, USA)A Hewlett-Packard Company (HP) HP48SX handheldcomputer workstation was interfaced to a SartoriusR200D semi-micro analytical balance via RS232C. Thehandheld computer functions as the front-panel (controlkeys and display) for the analytical balance. Handheldcomputer commands, to tare and send ASCII text fromthe analytical balance, are embedded within applicationspecific user programs. Application tasks are invoked viaprogram softkeys. Analytical balance acquired sampleweights, are sent to the handheld computer, where thereceived text, is simply stored, or is preprocessed and thenstored. With software available from HP, data can be sentfrom the handheld computer to IBM and IBM-compat-ible PCs and Apple Macintosh Computer workstations.

Handheld computer RS232 interfacing to a gaschromatograph mass spectrometer (GC/MS)workstation

Albert Marcus Morrishow and Wah Sang Lee (Mount SinaiSchool of Medicine, New York, NY, USA)A Hewlett-Packard Company (HP) HP48SX handheldcomputer was interfaced to a HP59970B Gas Chromato-graph/Mass Spectrometer (GC/MS) Chemstation

(workstation) via RS232. ASCII test of integrated GC/MS acquired data are sent from the GC/MS Chemstationto the HP48SX, where the received text is simply stored,or is preprocessed and then stored. With softwareavailable from HP, data can be sent from the HP48SX toIBM and IBM-compatible PCs and Apple MacintoshComputer workstations.

The use of a Zymark II robot to perform automatictitration of gastric juice samples

Mark Hamilton, Antonio Guglietta, Kim Collins, Mary Gerrelts,Lynne Hupe, Jim Fergus and Jim Marks (Warner-LambertParke-Davis, Pharmaceutical Research Division, 2800 PlymouthRoad, Ann Arbor, MI 48106-1047, USA)Gastric secretory studies are widely used in researchlaboratories to obtain data on gastrointestinal physiologyand pharmacology. Determination of hydrogen ionconcentration and gastric acid output requires basetitration of an aliquot of gastric juice to pH 7"0. Thisprocedure, when done manually, is very time consumingand only a limited number of samples can be handled atany given time.

This task has been automated utilizing a Zymate II robotoutfitted with a remote computer interface. The entireprocess is controlled by a master program that computesthe amount of NaOH to add based on output from a pHmeter. A comparison of results of titrations performedmanually and by the robot were almost identical.

Automated sample preparation for colorant analysiswith BenchMate workstation

Dean B. Higgins and Kathryn L. Jansen (Manufacturing QualityAssurance Organization, Eastman Kodak Company, Rochester,NY, USA)A Zymark BcnchMatc is being used to prepare colouredelectrographic toner samples for percentage colorantdetermination by spectrophotomctry. The coloured tonersamples must be dissolved in either N,N-dimethylacet-amide or concentrated sulfuric acid for this analysis. Useof the BenchMate for sample preparation minimizesanalyst contact by adding and mixing the solventsautomatically. Also, the total volume ofsolvent used (andwaste to be disposed of) can be reduced by a factor offive,from 100 ml to 20 ml per sample. Using BenchMatepreparation, no bias is observed and initial experimentsindicate reduced variability as compared to manualsample preparation. Optimization of the method andanalytical crossover results were described.

Use of Zymark BenchMate with Hewlett-Packarddiode array spectrophotometer for routine UV-VIS-NIR measurements

J. C. Hammontree and C. G. Zimba (Polaroid Corporation,Chemical Research Division, 750 Main Street 3D, Cambridge,MA 02139, USA)Absorption spectra of powders dissolved in variousorganic solvents are required in the authors’ laboratory.Using manual methods to prepare and measure the

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solutions, it typically takes 30-60 min for each sample. Atpresent, approximately three-quarters of a man-year isspent doing these analyses with the vast majority, 80-90%, of this time consumed in the preparation ofsolutions. The Zymark BenchMate allows the solutionsto be prepared under automated control and thendelivered, via a flow cell, to the spectrophotometer. Byreducing the time required to prepare solutions to 10% ofthe present commitment, the BenchMate will allow twoassociate scientists now committed to this work to beredirected to projects of a more appropriate skill level.Additionally, by allowing unattended overnight dataacquisitions, the BenchMate allows more rapid responsefor the routine service work that our laboratory performs.Furthermore, sample quantity requirements are reduced,lowering the amount of analyte required for the analysisand lowering the amount of waste organic generated.Progress toward integrating the Zymark BenchMate to aHewlett-Packard diode-array spectrophotometer forroutine UV-VIS-NIR absorption measurements werediscussed.

Capacitive proximity sensor aids robot navigation

Bill Mordan and John Shigeura (Applied Biosystems, FosterCity, CA, USA)

Fluid-handling laboratory robots require positional infor-mation regarding the liquid volumes that they work on, aswell as various solid objects such as trays and tubes. Thisposter described how a capacitive proximity sensorprovides this feedback in a robotic instrument thatexecutes DNA sequencing reactions. The software datastructures that use the feedback are illustrated.

Operating through a stainless steel pipette tip manipu-lated by the robot, the capacitive sensor enables the robotto locate the surface ofeach fluid volume in its workspace.This feedback reduces tip contamination and alerts theinstrument controller when pipetting failures haveoccurred. The sensor also allows the robot to locatedatum planes on the worksurface and on each well tray.With this information, the instrument controller cancompensate for errors in the tray and tip positions,thereby improving the accuracy of robot motions in thewells. Lastly, the sensor is used to identify well trays bydetecting the coded profile of each tray.

Automated assay for Vigabatrin and MDL 17,637 inSabril tablets using the Zymark BenchMate

W. Al Kentrup andJeffHuth (Analytical Development, MarionMerrell-Dow, 2110 E. Galbraith Road, Cincinnati, OH 45215,USA)

An automated method has been developed on theZymark BenchMate for the release/stability and contentuniformity assay of vigabatrin and its degradationproduct MDL 17,637 in sabril tablets. The BenchMatesamples an aliquot of the solubilized sabril tablet, filtersthe solution, performs the appropriate dilutions, andinjects an aliquot of the processed sample into thechromatograph. A Whatman Partsil-10 SCX column is

used to facilitate the separation of vigabatrin, MDL17,637, and excipient materials with UV detection at210 nm. The sample processing time is approximately10 min, and is equal to the chromatographic run time. Asa result, instrument lag-time is minimized.

Using the developed method, linearity studies for bothvigabatrin and MDL 17,637 produced correlation coeffi-cients of 1"000. Precision of the method was determinedboth chromatographically as well as using the gravi-metric function of the weigh station of the BenchMate.When ten aliquots of the same sample solution wereprocessed by the BenchMate, peak area counts forvigabatrin and MDL 17,637 had relative standarddeviations of 0’25% and 0"41% respectively. In sixseparate analytical runs (N 7-50 samples), 0’9 ml ofsolution sampled for dilution (’conc asp’) produced anaverage relative standard deviation of 0"27%. Similarly,the 2"25 ml of diluted solution dispensed (’sln disp’)produced an average relative deviation of 0"26%. Re-covery data for vigabatrin from spiked placebo samplesproduced an overall recovery of 100"4% (N 30). ForMDL 17,637, the average recovery from spiked placebosamples (N 30) over the range from 0’03% to 2"64%(w/w) MDL 17,637/active drug was 97"4%. For thehigher concentration range of 0" 11% to 11 "90% (w/w) an

average recovery of 98"8% was obtained (N 6).

The developed method has proven to be linear, accurateand precise. In addition, a cost reduction in labour andreagents has been realized using the automated methodover the conventional manual method.

Analysis of urinary catecholamines using Varian’sAccuCAT Column for sample preparation withZymark’s BenchMate workstation

Rachel A. Grace (Varian Sample Preparation Products HarborCity, CA, USA) and Lynn Jordan (Zymark Corporation,Hopkinton, MA 01748, USA)Catecholamines are neurotransmitters used by the bodyto regulate motor response and metabolism. Overpro-duction of catecholamines can cause hypertension orcongestive heart failure, and can also be indicative of theadrenal gland tumour known as pheochromocytoma.Quantitation of catecholamines in urine is used as an aidin diagnosis.

Catecholamines in the body are converted to an acidic orbasic metabolite by catechol-O-methylation. In the past,multiple solid phase extraction (SPE) technologies werenecessary to quantitate ’total catecholamines’. Newermethodologies use the concept of multimodal solid phaseextraction columns. The AccuCAT column is a mixedmode bed column for isolation of catecholamines, andallows one SPE cartridge to be used for determination ofcatecholamine levels.

With today’s trends towards automation, and theincreasing concerns about handling bodily fluids, thesample preparation and HPLC injection of catechol-amines was automated with the BenchMate Workstation.

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The patient samples were diluted and pH adjusted priorto placement on the BenchMate Workstation. TheBenchMate Workstation conditioned the AccuCAT col-umns, performed the SPE, diluted the eluant and injectedthe sample onto the HPLC.

This paper described the combination of two technol-ogies: the use of the AccuCAT columns and theBenchMate Workstation for automation of the Catechol-amine Analysis. The paper focused on the determinationof Vanillylmandelic Acid, the most common acidicmetabolite of catecholamines. The poster demonstratedthe transfer of the manual method to the BenchMateWorkstation. The automated method achieves goodlinearity and high percentage recoveries.

Aflatoxin determination using Vicam’s Aflatest-Pimmunoaffinity columns and Zymark’s BenchMateworkstation

Lynn Jordan (Zymark Corporation, Hopkinton, MA 01748,USA) and Kevin F. Donahue (Vicam L.P., Somerville, MA02145, USA)Aflatoxins are naturally occurring carcinogens producedby moulds found in grains, nuts and other commodities.Monitoring aflatoxin contamination is essential forproduction of quality food products and for compliancewith FDA guidelines.

This poster demonstrated the combination of two tech-nologies for improved aflatoxin analysis: Aflatest immu-noaffinit chromatography for aflatoxins and the Bench-Mate Workstation for automated sample processing. Theresult is increased sample throughput and highly repro-ducible data. Aflatest immunoaffinity columns are highlyselective to aflatoxin and permit rapid and efficientsample clean up. The affinity column clean up methoddoes not require the use of hazardous chemicals orlengthy sample preparation steps of some of the moretraditional aflatoxin methods. Aflatest columns utilizemonoclonal antibodies with a high specificity for aflatox-ins B 1, B2, G1 and G2. This method has received officialfirst action approval by the AOAC.

Combining the BenchMate Workstation with the Aflatestcolumns offer a number of additional benefits. With theBenchMate Workstation’s ability to precisely controlflow rates used to load, wash and elute columns, studieswere easily set up to optimize the method. An additionalfeature of the automated method is the ability of theworkstation to make direct HPLC injections, eliminatinganother sample handling step. Use of the BenchMateWorkstation to automate the aflatoxin analysis can meana time saving of 4 h a day with 30 analyses per day.

Data was presented from studies used to determine theoptimum loading flow rate for the Aflatest columns. Oncean optimized method was determined, linearity studiesand a comparison of the manual method to the Bench-Mate method was done. With the BenchMate automatedmethod, the data shows an improvement in the repro-ducibility, along with a time saving for the analyst. Theconsistency of the results achieved can lead to high levelsof confidence in producing a quality food product.

Automation of the assay for the determination ofenzyme activity of modified superoxide dismutaseinjectable drug product by the inhibition of cyto-chrome C reduction method

Charles Houck, Martin Echols, Timothy Dankanich and WilliamPickens (Sterling Research Group, Malvern, PA, USA)A robotic system has been developed which can conductunattended, the determination of enzyme activity ofpolyethylene glycol superoxide dismutase (PEG-SOD) inan injectable drug product. Modified superoxide dismu-tase is currently being investigated as a potential anti-inflammatory drug and antioxidant for reperfusion ofischaemic tissue.

The assay is based upon the method developed byMcCord and Fridovich. In this method, xanthine oxidaseacting aerobically upon xanthine generates superoxideradicals, this reaction can be followed at 550 nm bymonitoring the reduction of cytochrome c. PEG-SOD, bycompeting for the superoxide radicals, will inhibit thereduction of cytochrome c. The system produces highaccuracy and precision both interday and intraday.

A Sun 386i workstation controls a Hewlett-PackardWidmark robot, collects time based data from a Perkin-Elmer Lambda 2 spectrophotometer, processes the data,analyses the results and then makes appropriate decisionsconcerning the assay conditions based upon those results.This poster described the software developed, the hard-ware and how the Sun 386i workstation is being used as atest station controller to optimize the reaction conditionsand ensure the quality of the results generated.

Preparation of tablet samples for HPLC analysisusing a Zymate PySystem in a custom mode andsupplemented with a BenchMate workstation

B. A. Brown,J. E. Curley and L.J. Kostek (Pfizer, Inc., Groton,CT, USA)

The Pfizer Analytical Research & Development Robo-tic’s Laboratory has developed a diversified path toautomation oftablet sample preparation. Two automatedmethods have been developed for sample preparationusing a PyTechnology for totally automated testing or aZymark BenchMate for semi-automated preparations.These systems can be used separately or togetherdepending on the needs of the product.

A PySystem has been configured to perform a totallyautomated sample prep procedure. Custom programshave been written and integrated with some of theZymark Pysoftware to perform all the necessary steps ofsample preparation. This system will weigh a tablet andadd the dissolving solvent. The mixture is shaken and aportion ofthe extract is centrifuged. The clarified solutionis diluted and vortexed. The final solution is capped andstored for HPLC end analysis. The Zymark BenchMatemay be incorporated into this procedure if centrifugationproves to be inadequate and additional filtration isnecessary. The BenchMate has the capability of perform-ing filtration and dilution of a previously dissolved

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solution, which has been prepared either manually or bythe PyTechnology sample prep system.

This poster presented a detailed description of both theZymate sample prep system and the BenchMate pro-cedures. Validation experiments along with supportingdata were provided to indicate system suitability. Thesesystems are an alternative to manual preparation ofsamples for HPLC analysis.

The test is performed in 1000 ml of dissolution mediausing the USP XXII (basket) method I at 50 rpm and at37"0 + 0"5 C. Telescoping basket shafts are utilized toinitiate sample testing at specific time intervals.

The system is capable of performing four completedissolution tests in 24 h without operator intervention.This represents a 200% increase in productivity overconventional manual test methods.

Analysis of climate gases CO2, N20 and CH4 by widebore capillary gas chromatography

Bishal K. Sitaula, Luo Jiafa and Lars Bakken (AgriculturalUniversity of Norway, 1432 Aas-NLH, Norway)

A gas chromatograph method is described, which givessensitive and rapid (3"3 min) measurement of the threegreenhouse gases CO2, CH4 and N20. A wide borecapillary column operated at optimal conditions gave asufficient separation to allow a switch of the column flowbetween two detector systems (TCD and rID in series,and ECD) during the chromatogram, thus all three gasescould be analysed with a single sample injection.Application of a peristaltic pump allowed gas samples tobe transferred directly from gas sampling vials, through adrying agent (MgC103) and into the injection loop(0’2 ml). Back flushing of the injection system (pump anddrying agent) with He between each injection ensuredminimal carry over between samples (<0’6%). Experi-ence with storage of gas samples in vials with butylrubber septa was described.

Payback and benefits of a robotic dissolution system

Ronnie McDowell (Sandoz Pharmaceuticals, Q,A/MethodsDevelopment, East Hanover, NJ, USA)As part of the justification for the purchase of a roboticdissolution system it was determined that such a systemwould result in a net gain in productivity of 1’5 to 1"75analyst per year. Based upon this assumption, it wasfurther determined that full payback of such a systemcould be attained in less than two years. This posterpresented the authors’ experience with the system afterone year and included the initial installation, calibrationand validation of the system. Also presented were theresults ofa typical month ofautomated dissolution testingin our quality assurance laboratory. This data was usedto determine whether the forecasted payback period isattainable.

Automated dissolution testing using USP method I(baskets) and real-time HPLC analysis

D. W. Barrow and W. S. Conder (Bristol-Myers Squibb, P.O.Box 191, New Brunswick, NJ 08903-0191, USA)

A fully automated in vitro dissolution test for sustained-release capsules of various potencies has been achievedusing a Zymark Zymate I Plus robot (Zymark Corpor-ation, Hopkinton, MA) and on-line HPLC analysis.

Making robots smarter: a critical analysis of theforce feedback signals from the Zymate II robot andtheir use in error detection and correction

Naved A. Surve and Dennis S. France (Department ofLipids andLipoprotein Metabolism, Sandoz Research Institute, EastHanover, NJ 07936, USA)

Laboratory robots are being adapted for increasinglycomplex uses that requrie extreme precision. While theseadaptations might be crucial to the success of an assay,they also increase the chances of failure for the robot. Ifrobots were able to foresee conditions where they wereabout to crash, newer and more complex assays wouldbecome reliable. This would open the door for therobotization ofmore assays, freeing the time oflaboratoryworkers for more inventive tasks.

The Zymate II System provides feedback data on forcesexerted by the arm. By knowing what forces are exertedunder normal conditions and conditions of stress, it ispossible to create error checking programs which effec-tively give the robot the ability to recognize and correcterrors.

Both arm and hand force feedback parameters on twomicroplate management systems have been quantita-tively mapped. The limits, precision, reproducibility, andutility of these parameters in the context of roboticmicroplate management were presented. In addition,specific examples of error detection and correction werediscussed. A preliminary comparison was made betweenthe force feedback potentials of the Zymate II arm andthe faster Zymate XP arm.

Macintosh control of the Zymate robot: constructionof a multiple peptide synthesizer

Ronald N. Zuckermann, Michael A. Siani, Steven C. Banvilleand Janice M. Kerr (Chiton Corporation, 4560 Horton Street,Emeryville, CA 94608, USA)A multiple peptide synthesizer has been constructed byintegrating a Zymate robot arm and a peptide synthesisstation under the control of a Macintosh computer. TheMacintosh software co-ordinates the movements of theZymark robot arm (via a remote computer interface withthe Zymark controller), the switching of over 40 solenoidvalves and the monitoring of sensors (via an internaldigital I/O board). The Zymark hands are used to deliversolvent from pressurized spigot lines and to pipette aminoacid solutions from reservoirs to an array of reactionvessels. Liquid dispensing, reagent mixing and solvent

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removal are controlled from the Macintosh digital I/Oboard. The Macintosh was chosen because it permits afriendly user-interface and supports powerful program-ming languages. C language has been used to con-veniently accommodate peptide sequence, position andquantity information in multi-dimensional arrays whichare not supported by the EasyLab environment.

dictionary was then opened and edited by the System Vsoftware on the Mac. To enhance dictionary editing,Microsoft Ward v4.0 (Microsoft Corporation, Redmond,WA) was used in conjunction with the System V softwareto create and edit program text which was cut, copied,and pasted to and from the System V editor using theMacintosh’s clipboard.

Using a Macintosh IIx to perform off-line remoteprogramming

j. R. Ormand, A. E. Borgwardt, B. E. Kropscott and P. L.Morabito (The Dow Chemical Company, Health and Environ-mental Sciences, Midland, MI 48674, USA)A Macintosh IIx computer (Apple Computer, Inc.,Cupertino, CA), has been used in successful off-lineremote programming of a Zymate II System (ZymarkCorporation, Hopkinton, MA). Several applications havebeen edited using SoftPC EGA/AT v l.4 emulatingsoftware (Insignia Solutions, Inc., Sunnyvale, CA).

When Zymark introduced the System V controller, itprovided an interface between the robotic controller anda common computer environment, the IBM PC or PC-compatible computer. This interface offered the advan-tage ofhaving applications on the PC available to the userthat could not be offered by the Zymate controller.s.Applications such as word processing, and data manipu-lation software can now be used in conjunction with theSystem V software to enhance the user’s capabilities.Another advantage of a PC-based system is the con-venience of programming from a remote location andthen transferring the programs to the robotic system.

To accomplish remote editing on a Macintosh, theSystem V software was loaded onto the PC emulator harddrive. From the file conversion utility in the System Vmenu, the Zymate II dictionary (on a 5’25 in diskette)was converted to a binary .ZYD dictionary using an

Apple 5"25 in PC disk drive. The newly created .ZYD

To take this a step further, Zymark file conversionprograms were used to convert the binary .ZYD diction-ary to an editable text file (.ZYE dictionary) and viceversa. By opening the .ZYE text file in Microsoft Word,program text, positional coordinates, rack definitions(indexes and coordinates), and module commands wereedited on the Macintosh as a word processing document.Following editing, the .ZYE file was converted back to the.ZYD dictionary and finally back to the Zymate II 5"25 indisk.

An advantage to the dictionary conversion method ofediting is the ability to edit the entire dictionary as onedocument. This approach allows the user to edit in apowerful word processing environment instead of editingin the System V editor. Another advantage to dictionaryconversion is the ability to transfer positional code andrack definitions between dictionaries, which cannot beaccomplished with any of Zymark’s editors. The transferof programs also allows the user to create PyAppendabledisks for custom applications. A disadvantage ofediting a.ZYE file is that the format is more difficult to managethan the format in the System V editor. Also, whenediting the .ZYE file in a word processing application,care must be taken to insure that the application does notdelete or add control characters to the file. Anotherdisadvantage to this method of editing is that theinteraction with the System V software through the PCemulator is slow.

This poster described the methods used, as well as theadvantages and disadvantages of off-line remote pro-gramming on the Macintosh IIx.

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